PT1809273E - Use of 2-phenyl-1,2-ethanediol-(di)carbamates for treating epileptogenesis - Google Patents

Abstract

This invention is directed to methods for preventing, treating, reversing, inhibiting or arresting epilepsy and epileptogenesis in a subject comprising administering to the subject in need thereof a therapeutically effective amount of a compound selected from the group consisting of Formula (I) and Formula (II), or a pharmaceutically acceptable salt or ester thereof: wherein phenyl is substituted at X with one to five halogen atoms selected from the group consisting of fluorine, chlorine, bromine and iodine; and, R1, R2, R3, R4, R5 and R6 are independently selected from the group consisting of hydrogen and C1-C4 alkyl; wherein C1-C4 alkyl is optionally substituted with phenyl (wherein phenyl is optionally substituted with substituents independently selected from the group consisting of halogen, C1-C4 alkyl, C1-C4 alkoxy, amino, nitro and cyano).

Description

2-PHENYL-1,2-ETANODIOL (D1) CARBAMATES FOR TREATMENT OF EPILEPTOGENESE "

BACKGROUND OF THE INVENTION

FIELD OF THE INVENTION

This invention relates generally to the fields of pharmacology, neurology and psychiatry. In particular, the present invention provides compounds for use in the treatment, prevention, reversal, stopping or inhibition of epileptogenesis as defined in claim 1.

DESCRIPTION OF RELATED TECHNIQUE

Injuries or injuries of various types in the central nervous system (CNS) or peripheral nervous system (PNS) can produce profound and long lasting neurological and psychiatric symptoms and disorders. A common mechanism for producing these effects is the induction of convulsive activity or convulsion-like phenomena in the CNS or in the nerves and nodes of the PNS. Symptomatic of CNS paroxysmal disorders or electrical activity in the PNS, seizures or neurological mechanisms of convulsion are thought to underlie many of the pathological phenomena in a wide variety of neurological and psychiatric disorders. 1

A severe neurological condition characterized by seizures is epilepsy. Epilepsy is a common but devastating disorder that affects more than two and a half million people in the United States alone. Epilepsy describes a state in which a person has recurrent seizures due to an underlying, chronic process. Epilepsy refers to a clinical phenomenon and not to a single pathological entity, since there are many forms and causes of epilepsy. Using a definition of epilepsy as two or more unprovoked seizures, the incidence of epilepsy is estimated to be approximately 0.3 to 0.5 percent in different populations throughout the planet, with the prevalence of epilepsy estimated at 5 to 10 people for every 1000.

Based on the clinical and encephalographic phenomena, four subdivisions of epilepsy are recognized: epilepsy, large type mal (with subgroups: generalized, focal, jacksonian), small epilepsy type, psychomotor or temporal lobe epilepsy (with subgroups: own or tonic psychomotor (with flushing, pallor, tachycardia, hypertension, sweating or other visceral symptoms), and autonomic or diencephalic epilepsy (with flushing, pallor, tachycardia, hypertension, perspiration or other visceral symptoms).

Although epilepsy is one of the most advanced examples of a seizure disorder, a wide variety of neurological and psychiatric symptoms may have, like its etiology, seizures or related neurological phenomena such as seizures. In simple terms, a convulsion-related seizure or neurological phenomenon is a single discrete clinical event, caused by an excessive electrical discharge from a set of neurons or from a group of neurons 2 susceptible to convulsion through a process called " ictogenesis "; As such, the ictogenic seizures may be just the symptom of a disease. However, epilepsy and other convulsion-related analogous disorders are dynamic and often progressive diseases with a maturation process characterized by a complex and poorly understood sequence of pathological transformations. The development and maturation of such changes is the " epileptogenesis " process by which the broader collection of neurons than the normal brain is altered and subsequently becomes susceptible to abnormal, spontaneous, sudden, recurrent, excessive, i. and, seizures. The maturation of the epileptogenic process results in the development of " epileptogenic focus " according to which collections of abnormal discharge neurons or neurons susceptible to seizures form localized groups or " epileptogenic zones " dispersed along the cortical tissue. The epileptogenic zones are biochemically interconnected, so an abnormal ictogenic discharge is capable of cascading from zone to zone. As epileptogenesis progresses, the involved areas of the nervous system become more susceptible to a seizure and it becomes easier to trigger a seizure, resulting in progressively debilitating symptoms of seizure or seizure-related disorder.

Although ictogenesis and epileptogenesis may have a common origin in certain biochemical phenomena and neuronal pathways common in various diseases, the two processes are not identical. Itogenesis is the initiation and propagation of a seizure in a discrete time and space, a rapid and definite electrical / chemical event occurring over a period of time ranging from a few seconds to minutes.

Comparatively, epileptogenesis is a gradual biochemical or neuronal restructuring process whereby the normal brain is transformed by ectogenic events into an epileptogenically focused brain, having neuronal circuits that become sensitized and reactive to ictogenic events, making an individual increasingly susceptible to recurrence of spontaneous, episodic, time-limited seizures resulting in progressively debilitating seizure or convulsion-related symptoms and progressive tolerance to treatment. The maturation of an " epileptogenic focus " is a slow biochemical and / or structural process that usually occurs over months to years.

Epileptogenesis is a Two Phase Process: A " Phase 1 of epileptogenesis " is the initiation of the epileptogenic process prior to the first epileptic seizure or symptom of an analogous seizure related disorder and is often the result of some type of injury or trauma to the brain, heart attack, disease (e.g., infection such as meningitis) or trauma, such as an accidental blow to the head or a surgical procedure performed in the brain. A " Phase 2 epileptogenesis " refers to the process during which brain tissue which is already susceptible to epileptic seizures or phenomena related to seizure of a seizure-related analogue disorder becomes even more susceptible to seizures of increasing frequency and / or severity and / or severity becomes less reactive to the treatment.

Although the processes involved in epileptogenesis have not been definitively identified, some researchers believe that N-methyl-D-aspartate (NMDA) receptor-mediated regulation of excimer-coupled excitation is involved. Other researchers implicate the negative regulation of the inhibitory coupling between neurons, mediated by gamma-amino-butyric acid (GABA) receptors. In this process many other factors related to the presence, concentration or activity of NO (nitric oxide) or iron, calcium or zinc ions may be involved.

Although epileptic seizures are rarely fatal, large numbers of patients require medication to avoid the disruptive and potentially dangerous consequences of seizures. In many cases, the medication used to manage epileptic seizures or the symptoms of a seizure-related disorder is necessary for prolonged periods of time and in some cases the patient must continue to take the prescribed medication throughout life. In addition, such drugs are only effective in controlling symptoms and have side effects associated with chronic, prolonged use.

A wide variety of drugs available for the control of epileptic seizures include older agents such as phenytoin, valproate and carbamazepine (ion channel blockers), as well as newer agents such as felbamate, gabapentin, topiramate and tiagabine. In addition, for example, β-alanine has been reported to have anticonvulsive activity, NMDA inhibitory activity and GABAergic stimulatory activity but were not used to treat epilepsy.

Accepted drugs for the treatment of epilepsy are anticonvulsive agents or, more correctly designated, antiepileptic drugs (AEDs), wherein the term " antiepileptic " is synonymous with " anticonvulsant " or " anti-ictogenic ". These drugs therapeutically suppress seizures by blocking the onset of a single ictogenic event. But AEDs clinically available nowadays do not prevent the process of epileptogenesis.

In the treatment of seizures or related symptoms of seizure-related disorders, i.e. for diseases and disorders with neurological phenomena such as convulsions which may be apparently related to seizure disorders such as cyclic mood in Bipolar Disorder, impulsive behavior in patients with Disorders of Impulse Control or for convulsions resulting from brain injury, some AEDs may also be therapeutically useful. However, presently approved AEDs are incapable of prophylactically or therapeutically preventing the initial development or progressive maturation of epileptogenesis in an epileptogenic focus which also characterizes the seizure-related disorders. The poorly understood pathological mechanism underlying epileptogenesis certainly plays a role in the development of epilepsy and seizure-related disorders in a variety of clinical circumstances including spontaneous development or as a consequence of injury or trauma of many types in the central nervous system or peripheral. The current treatment of epilepsy is focused on suppression of convulsion by the administration of AED after the development of notorious clinical epilepsy. Although AEDs have positive effects on seizure suppression, currently available ones have been universally unsuccessful in preventing epileptogenesis, i. in the initial development or progression and worsening of epilepsy and other convulsion-related diseases. Even pre-treatment with AED does not prevent the development of epilepsy after injury or trauma to the nervous system. In addition, if AED therapy is discontinued, seizures typically reappear and, in unfortunate cases, worsen over time. At present, there is no clinically available method for treating, preventing, reversing, stopping or inhibiting the onset and / or progression of epilepsy or other seizure disorders or the many convulsion-related analogue disorders.

Furthermore, it is also believed that similar neurological mechanisms corresponding to epileptogenesis may be involved in the development and development of many disorders related to seizures clinically analogous to epilepsy which do not appear to be conspicuously " epileptic ", such as initial development and progressive aggravation observed in the mature pathology condition of Bipolar Disorder, Impulse Control Disorders, Obsessive-Compulsive Disorders, Schizoaffective Disorders, Substance Abuse or Dependence Disorders and many other psychiatric and neurological disorders. 7

Thus, despite the large number of drugs available for the treatment of epilepsy (ie, by suppressing epileptic seizure, ie seizures associated with epileptic seizures) and other seizure-related analogue disorders, there are no generally recognized drugs for treatment , prevention, reversal, stopping or inhibition of the process underlying epileptogenesis which may be etiological in many devastating neurological and psychiatric disorders, such as epilepsy and seizure related disorders including Bipolar Disorder.

At present there are no known methods of inhibiting the epileptogenic process to prevent the development of epilepsy or other convulsion-related analogue disorders in patients who have yet to present clinical symptoms thereof but who are either unknowingly or at risk of developing the disease. disease. In addition, there are no known methods for preventing development or reversing the epileptogenesis process, thereby converting collections of neurons into an epileptogenic zone, which is the source or is susceptible or is capable of participating in the convulsive activity of nervous tissue which does not exhibit abnormal, spontaneous, sudden, recurring or excessive electrical discharges or is not susceptible or capable of such convulsive activity. Moreover, there are no approved or unapproved medications recognized as possessing antiepileptogenic properties, i. (AEGD) (See, Schmidt, D. and Rogawski, A., Epilepsy Research, 2002, 50, 71-78).

Thus, there is a great need to develop safe and effective drugs or AEDs and methods of treatment that effectively treat, prevent, prevent, inhibit and reverse epileptogenesis in seizure-related neurological and / or psychiatric disorders.

SUMMARY OF THE INVENTION

This invention relates to compounds for use in the treatment and / or prevention, stopping, inhibiting and reversing epileptogenesis in a patient at risk of developing a seizure disorder or an analogous seizure disorder.

This invention is based, in part, on the previously unknown property of the carbamate compounds of the invention. These compounds are effective AEDs and may suppress epileptic seizures and, in addition, are strongly antiepileptogenic and may prevent the initial development and maturation of pathological changes in the nervous system which allow the occurrence and / or dissemination of seizures and related phenomena and may be capable of reverse those changes. Thus, the carbamate compounds of the present invention as used herein are true antiepileptogenic drugs (AEGDs) and have properties which are clearly different and not possessed by any other currently approved AED medication.

The compounds used in the present invention are of Formula 1 or Formula 2:

or a pharmaceutically acceptable salt or ester form thereof, wherein R 1, R 2, R 3 and R 4 are independently hydrogen or C 1 -C 4 alkyl, wherein C 1 -C 4 alkyl is unsubstituted or substituted with phenyl and wherein phenyl is unsubstituted or substituted substituted with up to five substituents independently selected from halogen, C1 -C4 alkyl, C1 -C4 alkoxy, amino, wherein amino is optionally mono or disubstituted with C1 -C4 alkyl, nitro or cyano; and χ, X2, X3, X4 and X5 are independently hydrogen, fluoro, chloro, bromo or iodo. Embodiments of the present invention include a compound of Formula 1 or Formula 2, wherein X 1, X 2, X 3, X 4 and X 5 are independently selected from hydrogen, fluorine, chlorine, bromine or iodine.

In certain embodiments, X4, X2, X3, X4 and X5 are independently selected from hydrogen or chlorine. In other embodiments, X 4 is selected from fluoro, chloro, bromo or iodo. In another embodiment, X 1 is chloro, and X 2, X 3, X 4 and X 5 are hydrogen. In another embodiment, R 4, R 2, R 3 and R 4 are hydrogen. The present invention provides enantiomers of Formula 1 or Formula 2 for use in the treatment of epileptogenesis in a subject in need thereof as defined in claim 1.

In certain embodiments, a compound of Formula 1 or Formula 2 will be in the form of a single enantiomer thereof. In other embodiments, a compound of Formula 1 or Formula 2 will be in the form of an enantiomeric mixture, in which one enantiomer predominates over the other enantiomer.

In another aspect, an enantiomer predominates in a range of from about 90% or more. In another aspect, an enantiomer predominates in a range of from about 98% or more.

In a compound of Formula 1 or Formula 2, R1, R2, R3 and R4 may be independently selected from hydrogen or C1 -C4 alkyl; and X 4, X 2, X 3, X 4 and X 5 are independently selected from hydrogen, fluorine, chlorine, bromine or iodine.

Prior to prophylactic or therapeutic administration of the composition of the invention to the individual, a determination will be made whether the subject is at high risk of developing epilepsy or seizure related disorders.

In certain embodiments of the present invention, a prophylactically or therapeutically effective amount of a compound of Formula 1 or Formula 2 for the treatment of epileptogenesis is in a range from about 400 mg / day to about 3000 mg / day (about 5 mg / , 7 mg / kg / day to about 43.0 mg / kg / day in a 70 kg human). Thus, the compounds and pharmaceutical compositions of the invention may be administered at a dosage of from about 5.7 to about 43.0 mg / kg / day (400-3000 mg / day in a 70 kg human), preferably , from about 6.4 to about 35.7 mg / kg / day (450-2500 mg / day in a 70 kg human), more preferably from about 7.1 to about 28.6 mg / kg / day (500-2000 mg / day in a 70 kg human) or even more preferred, from about 7.9 to about 21.4 mg / kg / day (550-1500 mg / day in a 70 kg human), or most preferably from about 8.6 to about 17.1 mg / kg / day (600-1200 mg / day in a 70 kg human). However, the dosages may vary depending upon the individual characteristics and tolerances of the subject and the exact nature of the condition being treated.

In certain embodiments, a prophylactic or therapeutically effective amount of a pharmaceutical composition for preventing, treating, reversing, stopping or inhibiting epileptogenesis comprising one or more of the enantiomers of a compound of Formula 1 or Formula 2 includes a pharmaceutically acceptable salt or ester acceptable amount thereof, admixed with a pharmaceutically acceptable carrier or excipient, according to which such composition is administered to the individual in need of treatment with an AEGD. Pharmaceutical compositions comprising at least one compound having Formula 1 or Formula 2 and one or more pharmaceutically acceptable excipients are administered to a subject in need thereof.

In another aspect, the subject or patient will be identified as being at risk for developing epilepsy or a seizure-related analogue disorder at the time of administration and on this basis will be considered to be a patient in need of treatment with an AEGD. 12

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 is a graph showing the effects of increasing CT doses on the number of neurons in different areas of the hippocampus counted 14 days after LI-pyl. Values are expressed as the number of neuronal cell bodies in each area of interest ± S.E.M.

Figure 2 is a graph showing the effects of increasing CT doses on the number of neurons in different nuclei of the amygdala counted 14 days after lycopene SE. Values are expressed as the number of neuronal cell bodies in each area of interest ± S.E.M.

Figure 3 is a graph showing the effects of increasing CT doses on the number of neurons in different nuclei of the thalam counted 14 days after lycopene SE. Values are expressed as the number of neuronal cell bodies in each area of interest ± S.E.M.

Figure 4 is a graph showing the effects of increasing CT doses on the number of neurons in different areas of the cortex counted 14 days after lycopene SE. Values are expressed as the number of neuronal cell bodies in each area of interest ± S.E.M.

Figure 5 is a graph showing the effects of increasing doses of CT in the latency period for the first spontaneous seizure. Values are expressed as mean latency in days for each group ± S.E.M. 13

Figure 6 is a graph showing the effects of increasing CT doses on the frequency of video-recorded spontaneous seizures over a 4-week period. Values are expressed as the mean number of seizures ± S.E.M. The total represents the total number of seizures observed during the 4 weeks of video recording and the mean represents the average number of seizures per week. The Anova test demonstrated a treatment effect on the total number of seizures (p = 0.045) and the mean number of seizures per week (p = 0.045)

Figure 7 shows the total number of seizures recorded in video over four weeks plotted according to the latency period for the first spontaneous seizure (SL = short latency period, LL = long latency period). Values are expressed as the mean number of seizures for each subgroup ± S.E.M. The ANOVA test did not show any significant effect of the treatment.

Figure 8: shows the correlation between the latency period for the first spontaneous seizure and the total number of seizures observed during the next four weeks.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides 2-phenyl-1,2-ethanediol monocarbamates and dicarbamates for use in the treatment and / or prevention of epileptogenesis as defined in claim 1.

The Carbamate Compounds of the Invention a or

Representative carbamate compounds according to the present invention include those having Formula 1 Formula 2:

Formula 1 wherein:

15

R 1, R 2, R 3 and R 4 are independently hydrogen or C 1 -C 4 alkyl and X 1, X 2, X 3, X 4 and X 5 are independently hydrogen, fluoro, chloro, bromo or iodo. " C 1 -C 4 alkyl " as used herein refers to substituted or unsubstituted aliphatic hydrocarbons having from 1 to 4 carbon atoms. They are specifically included in the definition of " alkyl " the aliphatic hydrocarbons which are optionally substituted. In a preferred embodiment of the present invention, the C1-C4 alkyl is unsubstituted or substituted by phenyl. The term " phenyl " as used herein alone or as part of another group is defined as a substituted or unsubstituted aromatic hydrocarbon ring group having 6 carbon atoms. They are specifically included in the definition of " phenyl " the phenyl groups which are optionally substituted. For example, in a preferred embodiment of the present invention, the " phenyl " is unsubstituted or substituted by halogen, C1 -C4 alkyl, C1 -C4 alkoxy, amino, nitro or cyano.

In a preferred embodiment of the present invention, X 1 is fluoro, chloro, bromo or iodo and X 2, X 3, X 4 and X 5 are hydrogen.

In another preferred embodiment of the present invention, X 1, X 2, X 3, X 4 and X 5 are independently chlorine or hydrogen.

In another preferred embodiment of the present invention, R1, R2, R3 and R4 are all hydrogen. 16

It is understood that the substituents and substitution patterns in the compounds of the present invention may be selected by a person skilled in the art to provide compounds which are chemically stable and which can be readily synthesized by techniques known in the art, as well as by the methods provided herein.

Representative 2-phenyl-1,2-ethanediol monocarbamates and dicarbamates include, for example, the following compounds:

Formula 3

Formula 4 17

Formula 6

Formula 7 18

Q

Formula 8

Suitable methods for synthesizing and purifying the carbamate compounds including the carbamate enantiomers used in the methods of the present invention are well known to those skilled in the art. For example, pure enantiomeric forms and enantiomeric mixtures of 2-phenyl-1,2-ethanediol monocarbamates and dicarbamates are described in U.S. Patents Nos. 5,854,283, 5,689,888 and 6,103,759, the disclosures of which are hereby incorporated by reference in their entirety.

The compounds for use in the present invention include isolated enantiomers of Formula 1 or Formula 2.

In a preferred embodiment, a pharmaceutical composition comprising the isolated S-enantiomer of Formula 1 is used to treat epileptogenesis in a subject.

In another preferred embodiment, a pharmaceutical composition comprising the isolated R-enantiomer of Formula 2 is used to treat epileptogenesis in a subject. 19

In another embodiment, a pharmaceutical composition comprising the isolated S-enantiomer of Formula 1 and the isolated R-enantiomer of Formula 2 may be used to treat epileptogenesis in a subject.

The compounds for use in the present invention also include the use of mixtures of enantiomers of Formula 1 or Formula 2. In one aspect of the present invention, an enantiomer will predominate. An enantiomer that predominates in the blend is one that is present in the blend in an amount greater than any of the other enantiomers present in the blend, e.g. in an amount greater than 50%. In one aspect, an enantiomer will predominate in a content of 90% or in a content of 91%, 92%, 93%, 94%, 95%, 96%, 97% or 98% or greater. In a preferred embodiment, the enantiomer that predominates in the composition comprising a compound of Formula 1 is the S-enantiomer of Formula 1. In another preferred embodiment, the enantiomer that predominates in the composition comprising a compound of Formula 2 is the R-enantiomer of Formula 2.

In one preferred embodiment of the present invention, the enantiomer which is present as the sole enantiomer or as the predominant enantiomer in a composition of the present invention is represented by Formula 3 or Formula 5, wherein X 1, X 2, X 3, X 4, X 5, R 1 , R 2, R 3 and R 4 are as defined above, or by Formula 7 or Formula 8. Formula 3

Formula 7 21

Q

Formula 8

An enantiomer of carbamate of Formula 1 or Formula 2 contains an asymmetric chiral carbon at the benzylic position, which is the aliphatic carbon adjacent to the phenyl ring.

An enantiomer which is isolated is one which is essentially free of the corresponding enantiomer. Thus, an isolated enantiomer refers to a compound which is separated via separation techniques or preparation free from the corresponding enantiomer. " essentially free " as used herein means that the compound is made up of a significantly higher proportion of an enantiomer. In preferred embodiments, the compound comprises at least about 90% by weight of a preferred enantiomer.

In other embodiments of the invention, the compound includes at least about 99% by weight of a preferred enantiomer. Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, including high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts or preferred enantiomers may be prepared by methods described herein. 22

Methods of preparing preferred enantiomers would be known to a person skilled in the art and are described, for example, in Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, S.H., et al., Tetrahedron 33: 2725 (1977); Eliel, E.L. Stereochemistry of Carbon Compounds (McGraw-Hill, NI, 1962); and Wilen, S.H. Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ. Of Notre Dame Press, Notre Dame, IN 1972).

Additionally, the compounds of the present invention may be prepared as described in U.S. Patent Number 3265728, 3313692 and U.S. Patents, previously referenced Nos. 5854283, 5698588 and 6103759. Treatment of Epileptogenesis

Using the compounds and compositions of this invention it is possible to suppress, control and prevent the process of epileptogenesis which results in aggravation, clinical progression or increased resistance to treatment of epilepsy and related seizure disorders or early de novo of these disorders and their symptoms as a consequence of some form of injury or trauma to the nervous system. The individual or patient in need of treatment is an individual who has not had the symptoms of epilepsy, i. seizures or seizures prior to the time of administration. The individual or patient will be identified as being at risk of developing epilepsy and on this basis will be considered to be a patient in need of treatment with an AEGD. 23

In suppressing the process of epileptogenesis the development of a convulsive disorder or a related disorder can be prevented in a subject who has some form of damage or deterioration of the nervous system or who is otherwise at risk.

Epilepsy The term epilepsy refers to a disorder of brain function characterized by the periodic and unpredictable occurrence of seizures (See, The Treatment of Epilepsy, Principles and Practice, Third Edition, Elaine Willie, MD, Editor, Lippincott Williams & Wilkins, 2001 ; Goodman & Gilman's The

Pharmacological Basis of Therapeutics, 9th edition, 1996). Convulsions that occur without obvious provocation are classified as epileptics. Epilepsy may be idiopathic or may be related to some type of injury, malformation or deterioration in the central nervous system at any stage of life. Typically an individual is considered to suffer from epilepsy after suffering two or more seizures that occur separate for more than 24 hours.

Clinically, an epileptic seizure results from a sudden and abnormal electrical discharge originating from a group of interconnected neurons in the brain or any other part of the nervous system. Depending on the type of epilepsy involved, the resulting activity of nerve cells may manifest itself by a wide variety of clinical symptoms, such as uncontrollable motor movements, changes in the patient's level of consciousness and the like. Epilepsy and epileptic seizures and syndromes can be classified in several ways (See, The

Treatment of Epilepsy, Principles & Practice, Third Edition, Elaine Willie, M.D. Editor, Lippincott Williams & Wilkins, 2001). However, as used herein the terms; " epilepsy ", " epileptic seizures " and " epilepsy syndromes " are intended to include all known types of epileptic seizures and syndromes including partial convulsions including simple, complex and partial seizures with development for qeneralized tonic-clonic seizures and generalized seizures, seizure convulsions and non-convulsive and unclassified seizures. The Epileptogenic Process The epileptogenic process generally consists of two phases. The first epileptogenic phase is known as the offending phase or initial lesion. The offense or initial injury is generally a damaging brain injury caused by one or more of a number of possible factors including, for example, traumatic brain injury, including blunt and penetrating head trauma or a neurosurgical procedure; infection of the CNS, such as, for example, bacterial meningitis, viral encephalitis, bacterial brain abscess or neurocysticercosis); cerebrovascular disease (such as heart attack or brain tumor including, for example, malignant gliomas, neurosurgery (such as, for example, craniotomies) and status epilepticus.

In some cases, the initial offense will be a consequence of developmental problems before birth (such as but not limited to asphyxia at birth, intracranial trauma at birth, metabolic disorders or congenital malformations of the brain) or a consequence of genetic determinants . The second epileptogenic phase is known as the latency phase. The second epileptogenic phase further includes the neural restructuring process, which is characterized by recurrent seizures (e.g., symptomatic epilepsy) or by the symptoms manifested in the seizure-related disorders. The epileptogenic process can also be observed among people who actually suffer from epilepsy or similar disorders related to seizures. Seizures experienced by persons suffering from epilepsy are themselves epileptogenic in that they tend to make subsequent seizures more likely or to widen the area of nerve tissue that is subject to seizure activity or to make the seizure disorder more resistant to seizures. treatment. The consequences of this process, for a patient having a seizure disorder, is that convulsions tend to become more frequent and more severe and are often more resistant to treatment with conventional AEDs.

Similarly, the related seizure-like response in neurological or psychiatric disorders analogous to epilepsy may become increasingly severe over time or resistant to treatment as the disorder matures.

In certain embodiments, phase 1 epileptogenesis may be initiated by factors other than those listed above, such as by ingestion of compounds with epileptogenic potential, e.g. psychotropic medications such as, for example, tricyclic antidepressants, clozapine and lithium, and the like. The compounds of the present invention are also intended to treat, prevent, stop, inhibit or reverse the development of epileptogenesis which has been initiated by factors that tend to increase the potential for an individual to become epileptogenic.

Therefore, in treating epileptogenesis, the compounds of the invention may prevent the development of seizures, in particular epileptic seizures. Accordingly, such compounds may be used to reduce the risk of developing epilepsy, halt the development of epilepsy (in particular the development of groups of neurons that are the source or are susceptible to ictogenic seizure), inhibit the development and maturation of epilepsy (in particular, the development of epileptogenic zones and the epileptogenic focus).

In addition, in treating, preventing, inhibiting, stopping or reversing epileptogenesis, according to the present invention, the development or progression of analogous neurological and / or psychiatric disorders whose etiology is based will be treated, prevented, inhibited, stopped or reversed. partially or totally in a mechanism of action type convulsion.

In some embodiments, the compounds of the present invention will advantageously be used to treat a patient who does not suffer or is known to be suffering from a condition which is known in the art to be effectively treated with the presently known anticonvulsive or antiepileptic (EDA) medications. In such cases, the decision to use the methods and compounds of the present invention would be made on the basis of determination if the patient is a patient in need of treatment with an antiepileptogenic drug (AEGD) " as that term is defined above.

The compounds of the present invention are for administration to a patient who has not developed epilepsy or any type of seizure disorder or an analogous seizure related disorder but who may be at high risk of developing seizures or an analogous seizure related disorder due to injury or trauma to the nervous system, including but not limited to cranial injury or heart attack or that may occur in the future, including but not limited to planned neurosurgical procedures or due to a known biochemical or genetic predisposition or by the detection of a biomarker of one or more of these disorders.

Thus, compositions of the present invention are directed to the treatment of epileptogenesis in an individual who is at risk of developing epilepsy or a seizure-related disorder but does not have epilepsy or clinical evidence of seizures.

An individual who is at risk of developing epilepsy but does not have epilepsy or another seizure disorder may be an individual in whom epilepsy has not yet been diagnosed but who is at a greater risk than epilepsy in the general population. This "higher risk" can be determined by the recognition of any factor in an individual, or in his / her clinical family history, physical examination or test that is indicative of a higher than average risk for developing epilepsy. Therefore, this determination that a patient may be at " higher risk " by any available means can be used to determine whether the patient should be treated with the compounds of the present invention.

Patients who are at highest risk would also include, but would not be limited to, those who have not suffered from deterioration or damage to their central nervous system but who are highly likely to suffer such deterioration or injury due to their physical state or their environment. This would include, but would not be limited to patients with a past Transient Ischemic Attacks (TIA) or known stenosis of the carotid artery or simply known atherosclerosis well as patients close to undergoing a neurosurgical procedure. In addition, they could be prophylactically administered with compounds of the invention, individuals likely to suffer from neurological deterioration due to war or sports injury; this would include combat soldiers or athletes in violent contact sports, such as pugilism.

Accordingly, in an illustrative embodiment, those who may benefit from treatment with the compounds of this invention can be identified using accepted screening methods to determine risk factors associated with epileptogenesis, epilepsy or other seizure disorders.

A determination that an individual has, or may be at risk of developing, epilepsy, or another seizure disorder would also include, for example, a medical evaluation that includes a thorough clinical history, a physical examination, and a relevant set of blood tests. It may also include an electroencephalogram (EEG), computerized tomography (CT), magnetic resonance imaging (MRI), or positron emission tomography (PET). A determination of a higher risk of developing epilepsy may also be made through genetic testing, including obtaining gene expression profiles or proteomic techniques (See, Schmidt, D. Rogawski, MA Epilepsy Research 50; 71-78 (2002), p. and Loscher, W, Schmidt D. Epilepsy Research 50: 3-16 (2002)).

These screening methods include, for example, conventional medical procedures for determining risk factors that may be associated with epileptogenesis, including but not limited to, for example, head trauma, closed or penetrating trauma, neurosurgical procedures, CNS infections, Trigeminal Neuralgia , bacterial or viral, cerebrovascular disease, including but not limited to heart attack or a past TIA, brain tumors, cerebral edema, cysticercosis, porphyria, metabolic encephalopathy, substance withdrawal including but not limited to withdrawal of sedative hypnotics or alcohol , abnormal perinatal past including anoxia in childbirth or injury of any kind at birth, cerebral palsy, learning disabilities, hyperactivity, past seizures, past epileptic status, family history of epilepsy, or any of a seizure disorder, inflammatory disease of the brain or including but not limited to cocaine and methamphetamine toxicity, parental consanguinity, and treatment with medications that lower the seizure threshold, including psychotropic medications such as antidepressant or antipsychotic medications . 30

In some embodiments, the compounds of the present invention would be used for the manufacture of a medicament for the purpose of treating a patient in need of treatment with an antiepileptogenic drug (AEGD). This would include the manufacture of a medicament for the purpose of treating a patient who is at risk of developing; epilepsy, a seizure disorder or seizure-related neurological phenomena related to epilepsy or seizure-related disorder as defined above, or any disorder in which the current clinical condition or prognosis of the patient could benefit from the suppression or inhibition of the epileptogenesis process to prevent the extension , aggravation or greater resistance to the treatment of any neurological or psychiatric disorder. Determining which patients who may benefit from treatment with an AEGD in patients who do not have clinical signs or symptoms of epilepsy or other seizure disorder may rely on a variety of " substitute " markers " or " biomarkers ". Such biomarkers would include but would not be limited to profiles of genes or proteins expression in tissue, blood or CSF or the presence of genetic markers, such as SNPs.

As used herein, the terms " substitute label " and " biomarker " are used interchangeably and refer to any anatomical, biochemical, structural, electrical, genetic or chemical indicator or marker that can be safely correlated with the actual existence or future development of epilepsy or a seizure disorder. In some cases, brain imaging techniques such as computed tomography (CT), magnetic resonance imaging (MRI) or positron emission tomography (PET), or other neuroimaging techniques may be used to determine whether an individual is at risk to develop one of the above disorders.

Examples of suitable biomarkers for the methods of this invention include, but are not limited to, determination by MRI, CT or other imaging techniques, sclerosis, atrophy or loss of volume in the hippocampus or the presence of temporal mesial sclerosis (MTS) or anatomical pathology relevant; detection in the blood, serum or tissues of the patient of a molecular species, such as a protein or other biochemical biomarker, e.g. elevated levels of ciliary neurotrophic factor (CNTF) or elevated serum levels of a neuronal degradation product; or other evidence of substitutable markers or biomarkers that the patient is in need of treatment with an antiepileptogenic drug, e.g. an EEG suggestive of a seizure disorder, epilepsy-related seizure-related neurological disorder or seizure-related disorder.

It is anticipated that many more of these biomarkers will be developed in the future using a wide variety of detection techniques. It is intended that any marker or indicator of the existence or possible future development of a convulsive disorder or epilepsy may be used in this invention to determine the need for treatment with the compositions and methods of this invention.

In some embodiments of the present invention, carbamate compounds suitable for use in the practice of this invention will be administered alone or concomitantly with at least one or more other therapeutic compounds or agents, e.g. with other antiepileptic drugs, anticonvulsant drugs or neuroprotective drugs or electroconvulsive therapy (ECT). In these embodiments, the present invention provides compositions for treating, preventing or reversing epileptogenesis

As used herein, the term " concomitant administration " or " association administration " of a compound, therapeutic agent or drug known with a compound of the present invention means administration of the drug and one or more compounds at a time such that the drug known to the compound will have a therapeutic effect. In some cases, this therapeutic effect will be synergistic. Such concomitant administration may involve concurrent (i.e., concurrent), prior or subsequent administration of the drug with respect to the administration of a compound of the present invention. One skilled in the art would have no difficulty in determining the height, sequence and appropriate dosages of administration for particular drugs and compositions of the present invention. Said one or more other compounds or therapeutic agents may be selected from compounds having one or more of the following properties: antioxidant activity; NMDA receptor antagonist activity, increased inhibition of endogenous GABA; NO synthase inhibitory activity; iron fixing capacity, e.g. an iron chelating agent; calcium fixing capacity, e.g. a Ca (II) chelating agent; zinc fastening capacity, e.g. a Zn (II) chelating agent; the ability to effectively block the sodium or calcium ion channels or to open the channels of potassium or chloride ions in the CNS of a patient, including known AEDs or are therapeutic agents useful in the treatment of Substance Abuse and Dependence, including but not limited to methadone, 33 antipsychotics, naloxone disulfiram, bupropion, benzodiazepines, buspirone, antidepressants, or naltrexone.

In some preferred embodiments, one or more other therapeutic compounds or agents would antagonize NMDA receptors by binding to NMDA receptors (eg, by binding to the glycine binding site of the NMDA receptors) and / or the agent would increase inhibition of GABA by decreasing the uptake of glial GABA.

In addition, said one or more other compounds or therapeutic agents may be any agent known to suppress seizure activity, even though such a compound is not known to inhibit epileptogenesis. Such agents would include but would not be limited to any effective AED or anticonvulsant known to a person skilled in the art or will be found in the future, for example suitable agents include, but are not limited to; carbamazepine, clobazam, clonazepam, etosuximide, felbamate, gabapentin, lamothiazine, levetiracetam, oxcarbazepine, phenobarbital, phenytoin, pregabalin, primidone, retigabine, talampanel, tiagabine, topiramate, valproate, vigabatrin, zonisamide, benzodiazepines, barbiturates or sedative hypnotics.

Definitions

As used herein, the term " epileptogenesis " means the biochemical, genetic, histological or other structural or functional processes or changes that render the nervous tissue, including the central nervous system (CNS) susceptible to recurrent, spontaneous seizures. In addition, the term " epileptogenesis " is also used in a broader sense herein to refer to the changes and processes that contribute to the clinical progression observed in patients with epilepsy or other convulsive disorder or an analogous seizure related disorder including but not limited to worsening or progression of the disorder and its symptoms or the development of " drug resistance ", wherein the disorder becomes more difficult to treat as a consequence of neurobiological modifications resulting in reduced drug sensitivity or recruitment by the epileptogenesis process of non-seizure prone nervous tissue. The term " inhibition of epileptogenesis " as used herein, refers to the prevention, slowing, stopping or reversal of the epileptogenesis process. The term " antiepileptogenic agent or drug " (AEGD) as used herein refers to an agent which is capable of inhibiting epileptogenesis when the agent is administered to a subject in need thereof. The term " convulsive disorder " as used herein refers to a disorder in a subject in which the subject suffers from seizures, e.g. seizures due to epileptic seizure. Convulsive disorders include, but are not limited to epilepsy and non-epileptic seizures, e.g. seizures due to the administration of a seizure agent or toxin to the subject.

As used herein, the term " individual " or " sick " is a human who has not yet manifested the symptoms of epilepsy but who may be in a high-risk group. 35

As used herein, the term " an individual in need of treatment with an AEGD " would include an individual who does not have epilepsy but who may be in a high risk group for developing seizures or a seizure related disorder due to central nervous system (CNS) or peripheral (PNS) injury or trauma. An individual or patient is considered to be at high risk of developing such seizure-related seizures or disorders due to injury or trauma to the CNS or SNP due to a known biochemical or genetic predisposition to seizure-related epilepsy or analogous disorder, or because a biomarker or verified substituent label of one or more of these disorders has been detected. The term " an individual in need of treatment with an AEGD " would also include any individual whose physical state or prognosis could benefit from treatment with an AEGD. This would include, but would not be limited to, any individual identified as being at greater risk of developing epilepsy, a seizure disorder or seizure-related neurological phenomena related to epilepsy or seizure related disorder as defined above, due to any predisposing factor. Predisposing factors include, but are not limited to: injury or trauma to any type of CNS or PNS; infections of the CNS, e.g. meningitis or encephalitis; anoxia; heart attack, i. cerebrovascular accidents (CVA); autoimmune diseases affecting the CNS, e.g. lupus; labor injuries, e.g. g. , perinatal asphyxia; heart attack; therapeutic or diagnostic vascular surgical procedures, e.g. carotid endarterectomy or cerebral angiography; coronary bypass surgery; spinal cord injury; hypotension; CNS injury by emboli, hyper or hypoperfusion of the CNS; hypoxia 36 affecting the CNS; known genetic predisposition for disorders known to respond to AEGD; CNS compression lesions; brain tumors, e.g. glioblastomas; bleeding or bleeding in or surrounding the CNS, e.g. g. , intracerebral bleeds or subdural hematomas; cerebral edema; febrile seizures; hyperthermia; exposure to toxic or poisonous agents; substance intoxication, e.g. g. cocaine; past family of seizure disorders past epileptic status; current treatment with medications that lower the seizure threshold, e.g. g. , lithium carbonate, thorazine or clozapine; evidence of substitutable markers or biomarkers that the patient is in need of treatment with an antiepileptogenic drug, e.g. g. , MRI scanning showing hippocampal sclerosis or other CNS pathology, elevated levels of serum neuronal degradation products.

As used herein, unless otherwise indicated, the term " epilepsy " should mean any disorder in which an individual (preferably an adult human, child or infant) experiences one or more seizures and / or tremors. Suitable examples include, but are not limited to, epilepsy (including, but not limited to location-related epilepsies, generalized epilepsies, epilepsies with generalized and local seizures, and the like), seizures as a complication of a disease or condition (such as seizures associated with encephalopathy, phenylketonuria, juvenile Gaucher disease, Lundborg progressive myoclonic epilepsy, heart attack, head trauma, stress, hormonal changes, substance use or withdrawal, use or abstinence from alcohol, sleep deprivation and the like) and the like. The term is intended to refer to the clinical disorder 37 regardless of the type of seizure, seizure origin, seizure progression or underlying cause or etiology. The term " antiepileptic drug " (AED) will be used interchangeably with the term " anticonvulsant agent ", and as used herein, both terms refer to an agent capable of; treating, inhibiting or preventing convulsive activity or ictogenesis when the agent is administered to a subject or patient.

As used herein, the term " an individual in need of treatment with an AED " would include an individual known to have epilepsy disease or who had repeated seizures or convulsions or manifested the symptoms of an analogous seizure related disorder regardless of the etiology of these symptoms.

As used herein, " halogen " should be chlorine, bromine, fluorine and iodine.

As used herein, the term " alkyl " whether used alone or as part of a substituent group, includes straight and branched chains. For example, alkyl radicals include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl and the like. Unless otherwise indicated, " C 1-4 alkyl " means a carbon chain composition of 1-4 carbon atoms.

When a particular group is " substituted " (eg, alkyl, phenyl, aryl, heteroalkyl, heteroaryl), such a group may have one or more substituents, preferably from one to five substituents, more preferably from one to three substituents, Preferably, from one to two substituents, independently selected from the list of substituents.

With respect to the substituents, the term " independently " means that where more than one such substituent is possible, such substituents may be the same or different from each other.

To provide a more concise description, some of the quantitative expressions given herein are not qualified with " about. &Quot; It will be understood that the term " about " whether used explicitly or not, all quantities provided herein refer to the actual value indicated and also refer to the approximate value indicated which would reasonably be deduced on the basis of current skill in the art, including approximations due to experimental conditions and / or of measurement to such an indicated value.

The terms " individual " or " sick " are used interchangeably herein and as used herein refer to a human who has undergone treatment, observation or experimentation.

As used herein, the term " compositions " is intended to cover a product comprising the specified ingredients in the specified quantities as well as any product resulting directly or indirectly from associations of the specified ingredients in the specified quantities.

In cases where the compounds according to this invention have at least one chiral center, they may exist as enantiomers. In cases where the compounds have two or more chiral centers, they may additionally exist as 39 diastereomers. It will be understood that all such isomers and mixtures thereof are within the scope of the present invention. In addition, some of the crystalline forms of the compounds may exist as polymorphs and, as such, are included in the present invention. In addition, some of the compounds may form solvates with water (i.e., hydrates) or common organic solvents and such solvates are also included within the scope of this invention. The present invention includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs will be functional derivatives of compounds which are readily converted in vivo to the required compound. Thus, in the methods of treatment of the present invention, the term " administration " will encompass the treatment of the various disorders described with the specifically disclosed compositions or with a composition which may not be specifically disclosed but which converts to the specified compound in vivo upon administration to the patient. Conventional procedures for selection and preparation of suitable prodrug derivatives are described, for example, in " Design of Prodrugs ", ed. H. Bundgaard, Elsevier, 1985.

For use in medicine, the salts of the compounds of this invention refer to " pharmaceutically acceptable salts " not toxic. However, other salts may be useful in the preparation of compounds according to this invention or of their pharmaceutically acceptable salts. Suitable pharmaceutically acceptable salts of the compounds include acid addition salts which, for example, can be prepared by mixing a solution of the compound with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid , succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid.

Moreover, in cases where the compounds of the invention carry an acid moiety, suitable pharmaceutically acceptable salts thereof may include the alkali metal salts, e.g. sodium or potassium salts; alkaline earth metal salts, e.g. calcium or magnesium salts; and salts prepared with suitable organic ligands, e.g. quaternary ammonium salts. Thus, representative pharmaceutically acceptable salts include the following; benzylsulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycolylarsanilate, hexylresorcinate , hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, ammonium salt of N-methylglucamine, oleate, pamoate (embonate), palmitate, pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodete and valerate.

Representative acids and bases which may be used in the preparation of pharmaceutically acceptable salts include the following: acids; including acetic acid, 2,2-dichloroacetic acid, acylated amino acids, adipic acid, alginic acid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, (+) - camphoric acid, camphorsulfonic acid, (+) - (1S) -camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, hydroxy-ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid, D-glucuronic acid, L-glutamic acid, α-oxo-glutaric acid, glycolic acid, hydrobromic acid, hydrochloric acid, (+) - L-lactic acid, (±) -DL-lactic acid, lactobionic acid, maleic acid, (-) - L-malic acid, malonic acid, (±) -DL-mandelic acid , methanesulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, L acid salicylic acid, 4-amino salicylic acid, sebaic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+) - L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid and undecylenic acid; and bases; including ammonia, L-arginine, benzothamine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2- (diethylamino) ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, L-lysine , magnesium hydroxide, 4- (2-hydroxyethyl) -morpholine, piperazine, potassium hydroxide, 1- (2-hydroxyethyl) -pyrrolidine, secondary amine, sodium hydroxide, triethanolamine, tromethamine and zinc hydroxide. The term " treat " or " treatment ", as used herein, refers to actions that give rise to any evidence of success in preventing or ameliorating an injury, pathology, symptoms or condition, including any objective or subjective parameters such as weakening; remission; decrease of 42 symptoms or make the lesion, pathology or condition more bearable by the patient; slowing down in the rate of degeneration or decline; making the end point of degeneration less debilitating; or improving the individual's physical or mental well-being.

Thus, the term " treatment " or " treat " is intended to include any action that improves, prevents, reverses, stops or inhibits the pathological process of epileptogenesis, as that term is defined and used herein. Treatment or amelioration of symptoms may be based on objective or subjective parameters; including the results of a physical examination, neurological examination and / or psychiatric evaluation.

Therefore, the term " treat " or " treatment " includes administration of the compounds or agents of the present invention to treat, prevent, reverse, stop or inhibit the epileptogenesis process. The term " therapeutic effect " as used herein, refers to the treatment, inhibition, weakening, reversal or prevention of epileptogenesis, the effects or symptoms of epileptogenesis, or side effects of epileptogenesis in an individual. The term " a therapeutically effective amount " or " a therapeutically effective dose " are used interchangeably and, as used herein, means a sufficient amount or dose of one or more of the compounds or compositions of the invention to produce a therapeutic effect, as defined above, in a subject or patient in need thereof; treatment, inhibition, weakening, reversion or prevention of epileptogenesis, the effects or symptoms of epileptogenesis, or side effects of epileptogenesis. The range of doses required for these distinct therapeutic effects will differ according to the characteristics of the subject or patient and the exact nature of the condition being treated. The term " pharmaceutical dosage form " as that term is used herein will refer to a form of one or more of the compounds or compositions of this invention together with pharmaceutically acceptable excipients to provide a formulation suitable for administration to a individual. The form may be adapted for administration by any appropriate route including, but not limited to oral, immediate and delayed, intravenous (IV), transdermal, intramuscular, intraventricular or nasal delivery and may comprise; tablets, pills, capsules, semi-solids, powders, sustained-release formulations, solutions, suspensions, emulsions, syrups, elixirs, aerosols or any other suitable compositions.

Dosage Regimens The amount of the carbamate compound required to treat epileptogenesis is defined as a therapeutically or pharmaceutically effective amount or dose. To achieve this aim, the compounds or compositions of this invention must be used in the correct therapeutically effective amount or dose, as described below. The dosage schedule and the amounts effective for this use, i. the dose or dosage regimen will depend upon a variety of factors including the exact nature of the disease or injury, the physical condition, weight, age of the patient and the like. In calculating the dosage regimen for a patient, the mode of administration is also taken into account. The range of doses that are expected to be effective in producing an antiepileptogenic effect in humans in severe and acute clinical conditions which are analogous to the lithium-pilocarpine rat-based model in Example 2 are determined by comparing known effective doses and blood levels in mice and humans.

In humans, the pharmacokinetics of one of the compounds of the invention referred to herein as Test Compound (TC), i.e. Formula 7, is linear after single oral administration and repeated in healthy adult males. Blood levels in humans;

In toxicological studies in humans, oral administration of the Test Compound at various doses over 7 days yielded the following C max and AUC (0-24): 1) At 100 mg. b.i.d. (200 mg in 24 hours or 2.85 mg / kg / day in a 70 kg human) C max was 3.6 microgram / mL and AUC was 42.2 microgram-hour / mL; 2) To 250 mg. b.i.d (500 mg in 24 hours or 7.14 mg / kg / day in a 70 kg human) C max was 8.2 microgram / ml and AUC was 102.3 microgram-hours / ml; 3) To 500 mg. b.i.d. (1000 mg in 24 hours or 14.28 mg / kg / day in a 70 kg human) C max was 17.2 microgram / mL and AUC was 204.1 microgram-hours / mL; 4) To 750 mg. b.i.d. (1500 mg in 24 hours or 21.4 mg / kg / day in a 70 kg human) Cmax was 28.2 microgram / mL and the AUC was 322.7 microgram-hours / mL. Blood levels in rats;

In toxicological studies in rats, oral administration of Test Compound (TC) for 8 days produced the following C max and AUC: 1) At 30 mg / kg / day Cmax was 9.33 microgram / mL and AUC was 97.32 microgram-hours / mL; 2) At 100 mg / kg / day C max was 20.63 micrograms / mL and

AUC was 230.33 microgram-hours / mL. 3) At 300 mg / kg / day Cmax was 70.34 microgram / mL and

AUC was 525.95 microgram-hour / mL

The doses tested in rats for the antiepileptogenic effect in Example 2 ranged from 30 mg / kg / day to 120 mg / kg / day. The lowest dose tested in this Example, i.e. 30 mg / kg, produced some measurable protective effects while the lowest dose tested in Example 1 was 10 mg / kg / day and produced minimal or no protective effects (See Examples 1 and 2 below). In rats, doses as low as 30 mg / kg / day of Test Compound (TC) would be expected to yield levels in the blood of; C max of 9.33 micrograms / mL and an AUC of 97.32 microgram-hours / mL. In humans, such levels in the blood would be expected at doses from about 500 mg / day to about 600 mg / day or from about 7.1 to about 8.6 mg / kg / day in a 70 kg human.

However, in Examples 1 and 2 relatively high doses and blood levels were required because of the very acute and acute animal model that was used and the need to produce demonstrated dramatic and rapid antiepileptogenic effects. Furthermore, in this severe acute animal model the compound was administered after the event or traumatic injury, i. i.e., the induction of epileptic status by administration of Li-Pilocarpine. This type of post-injury model probably correlates with similar acute and severe situations in human patients including but not limited to the initiation of measurement after CNS injury has already occurred.

In these situations the dosages required for an antiepileptogenic effect are expected to outweigh those that would probably be required in less acute or severe circumstances or in chronic situations and especially where the medication is used prophylactically.

In the situation where the medication is used prophylactically in a primary prevention or pre-treatment paradigm, the doses and blood levels required to produce clinically important antiepileptogenic effects will be expected to be somewhat less than the human equivalent of the 30 mg / kg dose / day in Examples 2.

As such, in most cases, doses that are expected to be therapeutically effective in clinical practice would be lower than those identified in this severe animal model of epileptogenesis. The ED 50 for the Seizure Prevention Test Compound in rats is from about 4 mg / kg to about 30 mg / kg (depending on the time and type of experience) whereby a minimum effective dose of 30 mg / kg in the models of epileptogenesis in rat is not unexpected. Based on these data, an expected human effective antiepileptogenic dose would be higher than the minimum dose required for anticonvulsive efficacy in humans. In a primary prevention paradigm, where administration is made well before any offense or pathological process is initiated, effective doses and blood levels in humans will be expected to be somewhat less than the human equivalent of the 30 mg / kg determined to be minimally effective in the rat lithium-pilocarpine model in Examples 1 and 2.

In human patients, in a primary prevention paradigm in which the medication would commence prior to any injury or deterioration of the human nervous system, the lower limits of an antiepileptogenic dose will be expected to be from about 400 mg / day to about 500 mg / day or about 5.7 mg / kg / day to about 7.14 mg / kg / day. In situations where the medication is initiated after the injury has been continued, the dose range will be expected to be somewhat higher, for example about 500 mg / day to about 600 mg / day or about 7.14 to about 8.6 mg / kg / day in a 70 kg human.

The compounds and compositions of the invention do not have a theoretical upper end with respect to their clinically effective dose range. Thus, the upper end of the therapeutically effective range would be determined by the maximum amount that could be tolerated by the patient. However, it would be expected, based on the data above that the highest dose, tested in rats, i. 120 mg / kg, which showed marked neuroprotective and antiepileptogenic effects, had a C max and AUC similar to or less than that produced in humans at a dose of 750 mg twice daily (1500 mg / day or approximately 21.4 mg / kg / day). This dose was easily tolerated in humans and the maximum tolerated dose would be considerably greater than that for many patients, possibly 2500 to 3000 mg / day or about 35.7 mg / kg / day to about 42.9 mg / kg / day in a human of 70 kg.

Thus, the compounds and pharmaceutical compositions of the invention may be administered at a dosage of from about 5.7 mg / kg / day to about 43.0 mg / kg / day (400-3000 mg / day in a 70 kg human) , preferably from about 6.4 to about 35.7 mg / kg / day (4550-2500 mg / day in a 70 kg human), more preferably from about 7.1 to (500-2000 mg / day in a 70 kg human), or even more preferably from about 7.8 to about 21.4 mg / kg / day ( 550-1500 mg / day in a 70 kg human), or most preferably from about 8.6 to about 17.1 mg / kg / day (600-1200 mg / day in a 70 kg human) . However, these dosages may be modified depending upon the individual characteristics and tolerances of the subject and the exact nature of the condition being treated.

Based on this disclosure, one of ordinary skill in the art will be able, without undue experimentation, in the light of such skill, to determine a therapeutically effective dose or amount of a particular substituted carbamate compound of the invention, to treat epilepsy and to produce a clinically effective antiepileptogenic effect significant. (See, e.g., Lieberman, Pharmaceutical Dosage Form (Vols. 1-3, 1992); Lloyd, 49 1999, The art, Science and Technology of Pharmaceutical Compounding, and Pickar, 1999, Dosage Calculations).

A therapeutically effective dose is also one in which any toxic or detrimental side effects of the active agent are more than compensated in clinical terms for the therapeutically beneficial effects. It should also be noted that for each particular individual, specific dosage regimens should be evaluated and adjusted over time according to the needs of the individual and professional judgment of the person administering or supervising the administration of the compounds. It will also be expected that the compositions of this invention may be initiated at a low or moderate dose and then increased to a fully therapeutically effective dose and blood level over a period of time.

For purposes of treatment, the compositions or compounds disclosed herein may be administered to the subject in a single bolus " administration, via continuous administration over an extended period of time or in a repeated administration protocol (eg, by a protocol of repeated administration hourly, daily or weekly). The pharmaceutical formulations of the present invention may be administered, for example, once or more times a day, 3 times a week or weekly. In one embodiment of the present invention, the pharmaceutical formulations of the present invention are administered orally once or twice daily.

In some embodiments, a regimen of treatment with the compounds of the present invention may begin, for example, after an individual experiences a brain injury or other initial offense, but before epilepsy is diagnosed in the subject, e.g. before the subject has a first or second seizure. In one embodiment, an individual being treated with a compound having epileptogenic potential, e.g. psychotropic drug, or an individual having a disease associated with a risk of developing epilepsy, e.g. autism, a treatment regimen with a carbamate compound of the present invention may be started.

In still other embodiments, a treatment regimen with the compounds of the present invention may commence before any deterioration or damage to the nervous system has occurred but at a time when such deterioration or injury may be expected or is likely to occur. For example, such a treatment regimen may begin before an individual undergoes a neurosurgical procedure or is likely to suffer head or brain trauma or other trauma, e.g. combat, violent or racing sports, recurring strokes, TIA, etc.

In certain embodiments; the carbamate compounds may be administered daily for a set period of time (week, month, year) after the occurrence of the brain injury or initial offense. An attending physician will know how to determine whether a carbamate compound has reached a therapeutically effective level, e.g. e.g., clinical examination of a patient or measuring levels of drug in blood or cerebrospinal fluid. One skilled in the art would be able to determine the maximum tolerated dose by physical examination to determine the presence and severity of side effects, such as unclear speech, lethargy, or reduced coordination. 51

In this context, a therapeutically effective dosage of the biologically active agent (s) may include repeated doses in a sustained treatment regimen which will produce clinically significant results to prevent, reverse, stop or inhibit epileptogenesis. Determination of effective dosages in this context is typically based on animal model studies followed by human clinical trials and is guided by determination of effective dosages and administration protocols that significantly reduce the occurrence or severity of symptoms or conditions of selective exposure in the subject. Suitable models in this regard include, for example, murine models, in rats, swine, felines, non-human primates and other accepted animal models known in the art. Alternatively, effective dosages may be determined using in vitro models (e.g., immunological and histopathological assays). Using such models, typical calculations and adjustments are typically necessary to determine a concentration and dose suitable for administering a therapeutically effective amount of the biologically active agent (s) (eg, amounts effective intranasally, effective for transdermal routes, effective intravenously or effective intramuscularly to elicit a desired response).

In an illustrative embodiment of the present invention, unit dosage forms of the compounds are prepared for standard administration regimes. In this way, the composition can be easily subdivided into smaller doses under the guidance of the physician. For example, unit dosages may be transformed into packaged powders, vials or ampoules and, preferably, in capsule or tablet form. The active compound present in these unit dosage forms of the composition may be present in an amount of, for example, from about 25 mg to about 800 mg or, preferably, in unit dosage amounts of from about 100 mg. 50, 100, 200, 250, 400, 450, 500 and 600 mg of one or more of the active carbamate compounds of the invention for single or multiple daily administration according to the particular need of the patient.

Carbamate Compounds as Drugs: The present invention provides isolated enantiomeric and enantiomeric mixtures of Formula 1 and / or Formula 2 as pharmaceuticals. The carbamate compounds are formulated as drugs to treat epileptogenesis.

Pharmaceutical Compositions The present invention further comprises pharmaceutical compositions containing one or more compounds of Formula 1 or Formula 2 with a pharmaceutically acceptable carrier.

Pharmaceutical compositions containing one or more of the compounds of the invention described herein as the active ingredient may be prepared by infixing the compound or compounds with a pharmaceutical carrier in accordance with standard pharmaceutical formulation techniques. The carrier may take a wide variety of forms depending on the desired route of administration (e.g., oral, parenteral). Thus, for oral liquid preparations, such as suspensions, elixirs and solutions, suitable carriers and additives include water, glycols, oils, alcohols, flavorings, preservatives, stabilizers, dyes and the like; for oral solid preparations, such as powders, capsules and tablets, suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrants and the like. The solid oral preparations may also be coated with substances, such as sugars or enteric coated to modulate the main absorption site. For parenteral administration, the carrier will generally consist of sterile water and other ingredients may be added to increase solubility or storage. Injectable suspensions or solutions may also be prepared using aqueous vehicles together with appropriate additives.

To prepare the pharmaceutical compositions of this invention, one or more compounds of the present invention as the active ingredient are intimately admixed with a pharmaceutical carrier according to conventional techniques of pharmaceutical formulation, which carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g. oral or parenteral, such as intramuscular. In preparing the compositions in the oral dosage form any of the usual pharmaceutical media may be used. Thus, for liquid oral preparations, such as, for example, suspensions, elixirs and solutions, suitable carriers and additives include water, glycols, oils, alcohols, flavorings, preservatives, dyes and the like; for solid oral preparations such as, for example, powders, capsules, oval tablets, gelatin capsules and tablets, suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrants and the like. Because of their ease of administration, the tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously used.

If desired, the tablets may be sugar coated or enteric coated by standard techniques. For parenteral use, the carrier will generally comprise sterile water, although other ingredients, for example, may be included for purposes such as solubility or storage. Injectable suspensions may also be prepared, in which case liquid carriers, suitable suspending agents and the like may be used.

Here the pharmaceutical compositions will contain, per dosage unit, e.g. tablet, capsule, powder, injection, teaspoonful and the like, an amount of the active ingredient necessary to deliver an effective dose as described above. Here the pharmaceutical compositions will contain, per unit dosage unit, e.g. g. , tablet, capsule, powder, injection, suppository, teaspoonful and the like, from about 10 mg to about 1000 mg of one or more compounds of Formula 1 or Formula 2, and preferably unit doses from about 25 mg to about 800 mg, and more preferably, in unit doses of from about 50 mg, 100 mg, 250 mg, 400 mg, 450 mg, 500 mg and 600 mg.

The pharmaceutical compositions may be administered at a dosage of from about 5.7 mg / kg / day to about 43.0 mg / kg / day (400-3000 mg / day in a 70 kg human), preferably, from about 6.4 mg / kg / day to about 35.7 mg / kg / day (450-2500 mg / day in a 70 kg human), more preferably from about 7.1 mg / kg / day to about 28.6 mg / kg / day (500-2000 mg / day in a 70 kg human), or even more preferably from about 7.9 mg / kg / day to about (550-1500 mg / day in a 70 kg human), or most preferably from about 8.6 to about 17.1 mg / kg / day (600- 1200 mg / day in a 70 kg human). However, the dosages may be modified depending on the requirements of the patient, the severity of the condition being treated and the compound to be used.

Advantageously, the compounds of the present invention may be administered in a single daily dose or the total daily dosage may be administered in divided doses of two, three or four times a day. In addition, the compounds of the present invention may be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal patches well known to those skilled in the art. When administered in the form of a transdermal delivery system, administration of the dosage will of course continue rather than alternate throughout the dosing regimen.

Preferably, these compositions are in unit dosage forms, such as tablets, pills, capsules, powders, granules, sterile parenteral solutions, aerosols or spray formulations, drops, ampoules, self-injecting devices or suppositories; for oral, parenteral, intranasal, sublingual or rectal administration, or for administration by inhalation or insufflation. Alternatively, the composition may be presented in a form suitable for once-weekly or once-monthly administration; for example, an insoluble salt of the active compound, such as the decanoate salt, may be suitable to provide a deposition preparation for intramuscular injection. To prepare solid compositions, such as tablets, the main active ingredient is mixed with a pharmaceutical carrier, e.g. g. , conventional ingredients for the preparation of tablets such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, and other pharmaceutical diluents, e.g. g. water to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention or a pharmaceutically acceptable salt thereof. When referring to such preformulation compositions as homogenous, it is meant that the active ingredient is uniformly dispersed throughout the composition, whereby the composition can be readily subdivided into equally effective dosage forms such as tablets, pills and capsules. This solid preformulation composition is then subdivided into unit dosage forms of the type described above containing from 0.020 mg to about 800 mg of the active ingredient of the present invention.

The tablets or pills of the novel composition may be coated or otherwise formulated to provide a dosage form having the advantage of prolonged action. For example, the tablet or pill may comprise an inner dosage component and an outer dosage component, the latter being in the form of an envelope thereon. The two components may be separated by an enteric layer which serves to resist disintegration in the stomach and allows the inner component to pass intact into the duodenum or to be delayed release. A variety of materials may be used for such enteric layers or coatings, such materials including a number of polymeric acids having materials such as shellac, cetyl alcohol and cellulose acetate.

Liquid forms in which the novel compositions of the present invention may be incorporated for oral or by injection administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical carriers. Suitable dispersing or suspending agents for aqueous suspensions include synthetic and natural gums such as tragacanth, gum arabic, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone or gelatin.

For example, for oral administration in the form of a tablet or capsule, the active drug component may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Further, when desired or necessary, binders may also be incorporated into the mixture; lubricants, disintegrants and dyes. Suitable binders include, without restriction, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as gum arabic, sodium tragacanth or oleate, sodium stearate, magnesium, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methylcellulose, agar, bentonite, xanthan gum, and the like. 58

Liquid dosage forms in suitably flavored suspending or dispersing agents, such as synthetic and natural gums, for example, tragacanth, gum arabic, methylcellulose and the like. Suspensions and sterile solutions are desired for parenteral administration. Isotonic preparations are used, which generally contain suitable preservatives when intravenous administration is desired.

Optimal dosages to be administered can be readily determined by those skilled in the art and will vary with the particular compound used, the mode of administration, the potency of the preparation, the mode of administration and the advancement of the disease state. In addition, factors associated with the particular patient being treated, including age, weight, patient regimen and time of administration, will result in the need to adjust dosages.

One skilled in the art will recognize that in vivo and in vitro assays using suitable, known and generally accepted cellular and / or animal models allow predicting the ability of a test compound to treat or prevent a particular disorder.

One skilled in the art will further recognize that human clinical trials including human trials of dosage range and efficacy can be completed in healthy and / or suffering patients of a given disorder, according to methods well known in the clinical art and medical.

In general, the carbamate compounds of the present invention may be administered as pharmaceutical compositions by any method known in the art for delivering therapeutic drugs including oral, buccal, topical, systemic (eg, transdermal, intranasal or suppository) or parenteral (eg, intramuscular, subcutaneous or intravenous administration.) Administration of the compounds directly into the nervous system may include, for example, administration via intracerebral, intraventricular, intact-cerebro, intrathecal, intracisternal, intraspinal or peri-spinal administration via administration of intracranial needles or catheters or intravertebral with or without pump devices.

The compositions may take the form of tablets, pills, capsules, semi-solids, powders, sustained release formulations, solutions, suspensions, emulsions, syrups, elixirs, aerosols or any other suitable compositions; and comprise at least one compound of this invention in association with at least one pharmaceutically acceptable excipient. Suitable excipients are well known to those skilled in the art and these and methods of formulating compositions can be found in classical references such as Alfonso AR: Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton PA, 1985, the disclosure of which is hereby incorporated by reference. is hereby incorporated by reference in its entirety and for all intents and purposes. Suitable liquid carriers, especially for injectable solutions, include water, saline, dextrose aqueous solution and glycols.

The carbamate compounds may be provided as aqueous suspensions. The aqueous suspensions of the invention may contain a carbamate compound mixed with excipients suitable for the manufacture of aqueous suspensions. Such excipients may include, for example, a suspending agent, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum arabic, and dispersing or wetting agents such as natural phosphatide (eg, lecithin), a condensation product of an alkylene oxide with a fatty acid (eg, polyoxyethylene stearate), a condensation product of ethylene oxide with a long chain aliphatic alcohol (eg, heptadecaethylene oxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol (eg, polyoxyethylene sorbitol monooleate) or a condensation product of ethylene oxide with a partial ester derived from fatty acid and a hexitol (eg, polyoxyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more colorants, one or more flavorants and one or more sweeteners, such as sucrose, aspartame or saccharin. The formulations may be adjusted for osmolarity.

Oily suspensions for use in the present methods may be formulated by suspending a carbamate compound in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil, such as liquid paraffin; or a mixture thereof. The oily suspensions may contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweeteners may be added to provide a palatable oral preparation, such as glycerol, sorbitol or sucrose. These formulations may be preserved by the addition of an antioxidant such as ascorbic acid. As an example of an oily vehicle for injection, see Minto, J. 61

Pharmacol. Exp. Ther. 281: 93-102, 1997. The pharmaceutical formulations of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil or a mineral oil, described above or a mixture thereof.

Suitable emulsifying agents include natural gums such as gum arabic and tragacanth gum, natural phosphatides such as soy lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan mono-oleate and products of condensation of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavorings, such as in the formulation of syrups and elixirs. Such formulations may also contain a demulcent, a preservative or a dye. The compound of choice, alone or in combination with other suitable components may be transformed into aerosol formulations (i.e., they may be " nebulized ") to be administered via inhalation. Aerosol formulations may be placed in pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen and the like.

Formulations of the present invention suitable for parenteral administration, such as, for example, intra-articular (intravenous), intravenous, intramuscular, intradermal, intraperitoneal, intraventricular and subcutaneous routes, may include sterile, aqueous or non-aqueous isotonic injections, which may contain antioxidants, buffers, bacteriostats and solutes to render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions which may include suspending agents, solubilizers, thickening agents, stabilizers and preservatives. Among the acceptable carriers and solvents which may be used are water and Ringers' solution, an isotonic sodium chloride. In addition, sterile fixed oils may be conventionally used as a solvent or suspending medium. For this purpose any mild fixed oil may be used including synthetic mono- or diglycerides. In addition, fatty acids, such as oleic acid, may also be used in the preparation of injectables. These solutions are sterile and generally free of undesirable matter.

In cases where the compounds are sufficiently soluble they can be directly dissolved in normal saline with or without the use of suitable organic solvents, such as propylene glycol or polyethylene glycol. Dispersions of the finely divided compounds can be prepared in aqueous sodium starch or carboxymethylcellulose solution, or in a suitable oil, such as peanut oil. These formulations may be sterilized by conventional, well known sterilization techniques. The formulations may contain pharmaceutically acceptable auxiliary substances as required to approximate the physiological states, such as pH adjusting agents and buffers, toxicity adjusting agents, e.g. g. , sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. The concentration of a carbamate compound in these formulations may vary greatly and will be primarily selected on the basis of volumes, fluid viscosities, body weight and the like, according to the particular mode of administration selected and the needs of the patient. For IV administration, the formulation may be a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, such as a 1,3-butanediol solution.

These formulations may be presented in unit dose or multidose sealed containers, such as ampoules and vials. Injectable solutions and suspensions may be prepared from powders, granules and sterile tablets of the type previously described.

A carbamate compound suitable for use in the practice of this invention may be and is preferably administered orally. The amount of a compound of the present invention in the composition may vary widely depending on the type of composition, unit dosage size, type of excipients and other factors well known to those of ordinary skill in the art. In general, the final composition may comprise, for example, from 1.0 wt% (wt%) to 90 wt% of the carbamate compound, preferably 10 wt% to 75 wt%, with the remainder being corresponding to the excipient or excipients.

Pharmaceutical formulations for oral administration may be formulated using pharmaceutically acceptable carriers well known in the art in dosages suitable for oral administration. Such carriers enable the pharmaceutical formulations to be formulated into unit dosage forms such as tablets, pills, powders, dragees, capsules, liquids, pellets, gels, syrups, pastes, suspensions, and the like. suitable for ingestion by the patient.

Formulations suitable for oral administration may consist of (a) liquid solutions, such as an effective amount of the packaged nucleic acid suspended in diluents, such as water, saline or PEG 400; (b) capsules, sachets or tablets, each containing a predetermined amount of the active ingredient, such as liquids, solids, granulates or gelatin; (c) suspensions in an appropriate liquid; and (d) suitable emulsions.

Pharmaceutical preparations for oral use may be obtained by combining the compounds of the present invention with a solid excipient, optionally grinding the resulting mixture and processing the granule mixture, after adding additional suitable compounds, if desired, to obtain tablets or cores of drageia. Suitable solid excipients are carbohydrate or protein fillers and include, but are not limited to, sugars, including lactose, sucrose, mannitol or sorbitol; corn starch, wheat, rice, potatoes or other plants; cellulose, such as methylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose or sodium carboxymethylcellulose; and gums including gum arabic and tragacanth; as well as proteins, such as gelatin and collagen.

If desired, disintegrants or solubilizers, such as crosslinked polyvinylpyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate may be added. The tablet forms may include one or more of lactose, sucrose, mannitol, sorbitol, calcium phosphates, corn starch, potato starch, microcrystalline cellulose, gelatin, colloidal silicon dioxide, talc, magnesium stearate, stearic acid, and other pharmaceutically compatible excipients, dyes, fillers, binders, diluents, buffers, humectants, preservatives, flavorings, dyes, disintegrants and vehicles.

The tablet forms may comprise the active ingredient in a flavor, e.g. sucrose, as well as pastilles comprising the active ingredient on an inert basis, such as gelatin and glycerin or emulsions of sucrose and gum arabic, gels and the like containing, in addition to the active ingredient, carriers known in the art.

The compounds of the present invention may also be administered in the form of suppositories for rectal administration of the drug. These formulations may be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at rectal temperatures and thereby will melt the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

The compounds of the present invention may also be administered by the intranasal, intraocular, intravaginal and intra rectal routes including suppositories, insufflation, powders and aerosol formulations (for examples of steroid inhalants, see Rohatagi, J. Clin. Pharmacol. 1193, 1955, Tjwa, Ann.

Allergy Asthma Immunol. 75: 107-111, 1995). creams,

The compounds of the present invention may be administered transdermally via a topical route, formulated as applicator bars, solutions, suspensions, emulsions, gels, ointments, pastes, jellies, paints, powders and aerosols.

Encapsulating materials may also be used with the compounds of the present invention and the term " composition " may include the active ingredient in association with an encapsulating material as a formulation, with or without other carriers. For example, the compounds of the present invention may also be administered as microspheres for slow release in the body. In one embodiment, the microspheres can be administered via intradermal injection of drug-containing microspheres (eg, mifepristone), which is released subcutaneously slowly (see Rao, J. Biomater Sci, Ed., 7: 623-645, 1995, as formulations of biodegradable and injectable gel (see, eg, Gao, Pharm Res. 12: 857-863, 1995), or as microspheres for oral administration (see, eg, Eiles, J. Pharm Pharmacol 49: 669-674, 1997) Both transdermal and intradermal routes provide for steady administration for weeks or months Hosts may also be used in the administration of the compounds of the present invention.

The compositions of this invention may be administered in a variety of oral dosage forms adapted for slow or controlled release. For example, the composition may be placed in an insoluble capsule with an orifice at one end and a swellable fluid absorbent composition in the capsule on the opposite side of the perforated end. Upon administration, the fluid absorbent composition absorbs water from the patient's GI apparatus and dilates and forces the active drug out through the orifice at a known and controllable rate. Many other delayed release or controlled release dosage forms known in the art may also be used in conjunction with the methods and compositions of this invention. 67

In another embodiment, the compounds of the present invention may be administered through the use of liposomes, which fuse with the cell membrane or undergo endocytosis, i. using ligands attached to the liposome that bind to the cell surface membrane protein receptors resulting in endocytosis. In using liposomes, in particular when the surface of the liposome carries specific ligands for target cells, or when they are otherwise preferentially targeted to a specific organ, the administration of the carbamate compound in target cells in vivo. (See, e.g., Al-Muhammed, J.

Microencapsul. 13: 293-306, 1996; Chonn, Curr. Opin. Biotechnol. 6: 698-708, 1995; Ostro, Am. J. Hosp. Pharm. 46: 1576-1587, 1989).

The pharmaceutical formulations of the invention may be provided as a salt and may be prepared with many acids, including but not limited to hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic, and the like acids. The salts tend to be more soluble in aqueous or other protonic solvents than the corresponding free base forms.

In other cases the preferred preparation may be a lyophilized powder which may contain, for example, any or all of the following: 1 mM-50 mM histidine, 0.1% -2% sucrose, 2% -7% mannitol, at a pH range of 4.5 to 5.5 which is combined with buffer prior to use.

The pharmaceutically acceptable salts and esters refer to salts and esters which are pharmaceutically acceptable and have the desired pharmacological properties. Such salts include salts which may be prepared when acidic protons are present in the compounds which are capable of reacting with inorganic or organic bases. Suitable inorganic salts include those prepared with the alkali metals, e.g. sodium and potassium, magnesium, calcium and aluminum. Suitable organic salts include those prepared with organic bases, such as amine bases, e.g. ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like.

Pharmaceutically acceptable salts may also include the acid addition salts prepared by reacting amine units in the parent compound with inorganic acids (eg hydrochloric and hydrobromic acids) and organic acids (eg acetic acid, citric acid, maleic acid and alkane- and arenesulfonic acids, such as methanesulfonic acid and benzenesulfonic acid). Pharmaceutically acceptable esters include the esters of carboxyl, sulfonyloxy and phosphonoxy groups present in the compounds. Where two acid groups are present, a pharmaceutically acceptable salt or ester may be a mono-acid mono-salt or ester or a salt or diester; and similarly when there are more than two acid groups present, some or all of the groups may be salified or esterified.

The compounds designated in this invention may be present in unsalted or non-esterified form, or in salified and / or esterified form, and the indication of such compounds is intended to include the parent compound (unsalted and non-esterified) and pharmaceutically acceptable salts and esters thereof acceptable. The present invention includes pharmaceutically acceptable salt and ester forms of Formula 1 and Formula 2. There may be more than one crystalline form of an enantiomer of Formula 1 or Formula 2 and as such are also included within the present invention.

A pharmaceutical composition of the invention may optionally contain, in addition to a carbamate compound, at least one other therapeutic agent useful in the treatment of a disease or condition associated with epilepsy or epileptogenesis or an analogous seizure related disorder

Methods of formulating pharmaceutical compositions have been described in a large number of publications such as Pharmaceutical Dosage Forms: Tablets. Second edition. Magazine and Extension. Volumes 1-3, edited by Lieberman et al; Pharmaceutical Dosage Forms: Parenteral Medications. Volumes 1-2, edited by Avis et al; and Pharmaceutical Dosage Forms: Disperse Systems. Volumes 1-2, edited by Lieberman et al; published by Mareei Dekker, Inc., the disclosures of which are hereby incorporated by reference in their entirety and for all intents and purposes.

The pharmaceutical compositions are generally formulated as sterile, substantially isotonic and in full compliance with all Good Manufacturing Practices (GMP) regulations of the U.S. Food and Drug Administration.

The following Examples are set forth to aid in the understanding of the invention and are not intended to and should not be construed as immitating in any manner the invention set forth in the claims which follow. 70

EXAMPLES The activity of an isolated S-enantiomer of Formula 1 (e.g., Formula 7), referred to herein as the " Test Compound " or " TC " or " test compound " was evaluated in the following experiments to determine the efficacy of the compound for neuroprotection and in the treatment of epileptogenesis in the lithium-induced temporal lobe epilepsy model and pilocarpine in the rat.

Example 1 The lithium-pilocarpine model of temporal lobe epilepsy The rat-induced model of lithium-associated pilocarpine (Li-Pyl) reproduces most of the clinical and neurophysiological specificities of temporal lobe epilepsy in humans (Turski et al., 1989, Synapse 3: 154-171; Gentleman, 1995, Ital J Neurol Sci., 16: 33-37). In adult rats, systemic administration of pilocarpine leads to epileptic status (SE). The lethality rate reaches 30-50% during the first few days. In surviving animals, neuronal deterioration predominates in hippocampal formation, in the piriform and entorhinal cortices, thalamus, amygdaloid complex, neocortex and substantia nigra. This acute seizure period is followed by a " silent " free of seizures that persists for an average duration of 14-25 days after which all animals exhibit recurrent spontaneous seizures at the usual frequency of 2 to 5 per week (Turski et al., 1989, Synapse 3: 154-171, Gentleman, 1995, Ital J Neurol Sci 16: 33-37; Dube et al., 2001, Exp Neurol 167: 227-241). 71 Lithium-pilocarpine and treatments with the test compound

Male Wistar rats weighing 225-250 g, supplied by Janvier Breeding Center (Le Genest-St-Iste, France) were housed under controlled current conditions (light / dark cycle, 7.00 am-7.00 p.m.) , with food and water available ad libitum. All animal experiments were carried out in accordance with the rules of the Council of the European Communities Directive of 24 November 1986 (86/609 / EEC) and the French Department of Agriculture (License No. 67-97). For electrode implantation, the rats were anesthetized by an i.p. 2.5 mg / kg diazepam (DZP, Valium, Roche, France) and 1 mg / kg ketamine hydrochloride (Imalgene 1000, Rhone Merrieux, France). Four simple contact recording electrodes were placed on the skull on the parietal cortex, two on each side.

Induction of status epilepticus:

Treatment with the test compound and occurrence of spontaneous recurrent seizures (SRS)

All rats received lithium chloride (3 meq / kg, i.p., Sigma, St. Louis, Mo, U.S.A.); about 20 h later, the animals were placed in boxes in pexiglas to register the cortical background EEG. Methylescopolamine bromide (1 mg / kg, s.c., Sigma) was administered to restrict the peripheral effects of the seizure. SE was induced by injecting pilocarpine hydrochloride (25 mg / kg, s.c., Sigma) 30 min after methylscopolamine. Bilateral EEG cortical activity was recorded during the total duration of SE and behavioral changes were recorded.

The effects of increasing doses of the Test Compound were studied in the 3 groups of rats. The animals in the first group received 10 mg / kg of the test compound, ip, 1 h after the appearance of SE (pilo-TClO), while the animals in groups 2 and 3 received 30 and 60 mg / kg of Test Compound pilo-TC30 and pilo-TC60), respectively.

Another group was injected with 2 mg / kg diazepam (DZP, i.m.) 1 h after the appearance of SE which is the standard treatment of the subject to improve the survival of the animals after SE (pilo-DZP). The control group received saline instead of pilocarpine and the Test Compound (saline-physiological saline). The pilo-Test Compound mice surviving the SE were then injected, about 10 h after the first injection of the test compound, with a second i.p. of the same dose of the test compound and were kept under a twice-daily treatment with the test compound for a further 6 days. Pilo-DZP received a second injection of 1 mg / kg DZP on SE day about 10 h after the first. Thereafter, the Pilo-DZP and physiological serum-saline rats received twice-daily equivalent volume of saline.

The effects of the test compound on the EEG and the latency period for the occurrence of SRS were investigated by daily recording of the animals on video for 10 h per day and by recording electrographic activity twice weekly for 8 h. 73

Quantification of cell densities Quantification of cell densities was performed 6 days after SE in 8 pilo-DZP rats, 8 pilo-TClO rats, 7 pilo-TC30 rats, 7 pilo-TC60 rats and 6 rats physiological saline-physiological serum. At 14 days after SE, the animals were deeply anesthetized with 1.8 g / kg of pentobarbital (Dolethal®, Vetoquinol, Lure, France) Brains were removed and frozen successive slices of 20 pm were cut in a cryostat, air dried The quantification of the cell densities was performed with a microscopic grid of 1 cm2 with 10 x 10 cells in coronal sections according to the stereotactic coordinates of the atlas of the rat brain (see Paxinos G, Watson C ( 1986) The Rat Brain in Stereotaxic Coordinates, 2nd Academic Press, San Diego)

The cell counts were performed blindly twice and averaged at least 3 values of 2 adjacent sections in each individual animal. The counts involved only cells greater than 10 μm, the smallest being considered as glial cells.

Revelation of Timm

At 2 months after the occurrence of spontaneous recurrent seizures the development of mossy fiber in the rats in the chronic period exposed to the test compound or DZP was examined and in 3 rats saline-physiological serum. The animals were deeply anesthetized and perfused transcardiacally with 74 physiological saline followed by 100 mL of 1.15% (w / v) Na 2 S in 0.1 M phosphate buffer and 100 mL of 4% (v / v) formaldehyde in phosphate buffer 0.1 M. The brains were removed from the skull, postfixed in 4% formaldehyde for 3-5 h, and sections were cut at 40 Âμm in a sliding vibratome and mounted on gelatin coated slides.

The following day the sections were grown in the dark in a solution at 25 ° C of 50% (w / v) gum arabic (160 mL), sodium citrate buffer (30 mL), 5.7% (w / v) hydroquinone (80 mL) and 10% (w / v) silver nitrate (2.5 mL) for 40-45 min. Sections were then washed with tap water at 40øC for at least 45 min, rinsed quickly with distilled water and allowed to dry. They were dehydrated with ethanol and covered with a blade. The development of mossy fiber was evaluated according to the criteria previously described in the dorsal hippocampus (Cavazos et al., 1991, J Neurosci 11: 2795-2803), which are as follows: 0 - no granules between the tips and the crown of the DG; 1 - scarce granules in the supragranular region in an irregular distribution between the tips and the crown of the DG; 2 - more numerous granules in a continuous distribution between the ends and the crown of the DG; 3 - prominent beads in a continuous pattern between the tips and the crown, with occasional portions of granules confluent between the tips and the crown; 4 - prominent beads forming a dense confluent web between the tips and the crown and dense confluent laminar web of beads extending into the inner molecular layer. 75

Data analysis

To compare the characteristics of the SE in pilo-saline and pilo-test animals, a non-paired Student's t-test was used. The comparison between the number of rats that suffered seizures in both groups was performed by a Chi square test. For neuronal deterioration, statistical analysis between groups was performed using ANOVA followed by Fischer's test for multiple comparisons using the Statview software (Fisher RA, 1946a, Statistical Methods for Research Workers (10th edition) Oliver & Boyd, Edinburgh, Fisher RA , 1946b, The Design of Experiments (4th edition) Oliver & Boyd, Edinburgh)

Behavioral and EEG characteristics of the epileptic state of lithium-pilocarpine

A total number of Sprague-Dawley rats weighing 250-330 g was subjected to Li-pyl induced SE. The behavioral characteristics of the SE were identical in the pilo-saline and pilo-compound assay groups. Within less than 5 min after injection of pilocarpine, the rats developed diarrhea, piloerection and other signs of cholinergic stimulation. During the next 15-20 min the rats had their heads tilted, scraped, chewed and had an exploratory behavior. Recurrent seizures began about 15-20 min after administration of pilocarpine. These seizures, which associated episodes of myoclonus of the head and bilateral of the hind limbs with steepness and fall, progressed to SE at about 35-40 min after pilocarpine, as previously described (Turski et al., 1983, Behav Brain Res 9: 315-335.). 76

EEG patterns during SE

During the first hour of SE, in the absence of pharmacological treatment, the amplitude of the EEG increased progressively as the frequency decreased. At less than 5 min after injection of pilocarpine, normal background EEG activity was replaced by rapid low-voltage activity in the cortex at the same time as theta rhythm (5-7 Hz) appeared in the hippocampus. At 15-20 min, a high tension fast activity overlapped the hippocampal theta rhythm and isolated high voltage peaks were recorded only in the hippocampus while the cortex activity did not change substantially.

At 35-40 min following injection of pilocarpine, the animals developed typical electrographic seizures with high tilt activity present in the hippocampus and cortex which occurred first as activity pulses preceding the seizures and were followed by continuous high voltage peak sequences and multiple peaks that lasted until the administration of DZP or the test compound. At about 3-4 h SE, the EEG of the hippocampus was characterized by periodic electrographic discharges (PED, about one / sec) in the pilo-DZP group and pilo-10 in the hippocampus and cortex. The amplitude of background EEG activity was low in pilo-TC60 animals. At about 6-7 h of SE the activity of electrical peaks in the cortex and hippocampus in the DZP and TC10 treated rats still remained as the EEG amplitude decreased and returned to background levels in the hippocampus of TC30 rats and in both structures in TC60-treated rats. There was no difference between groups TC10, TC30 and TC60. At about 9 h of SE, isolated peaks were recorded in the hippocampus of rats treated with test compound and occasionally in the cortex. In both structures, background activity was of very low amplitude at that time.

Mortality induced by SE

During the first 46 h after SE, the degree of mortality was similar in pilo-DZP (23%, 5/22), pilo-TClO (26%, 6/23) and pilo-TC30 rats (20%, 5 / 25). The mortality rate was largely reduced in pilo-TC60 rats, in which it reached only 4% (1/23). The difference was statistically significant (P < 0.01).

Characteristics of the silent phase EEG and occurrence of spontaneous recurrent seizures

EEG patterns during the silent period were similar in the pilo-DZP and pilo-TClO, 30 or 60 rats. At 24 and 48 h days after SE, background EEG was further characterized by the occurrence of PEDs in which overlapping waves or peaks. Between 1 and 8 h after injection of the test compound or vehicle injection, there was no change in the pilo-DZP or pilo-TClO groups. In the TC30 and TC60 rats, the frequency and amplitude of the PEDs decreased as fast as 10 min after injection and were replaced by large amplitude peaks in the TC30 group and small amplitude in the TC60 group. At about 4 h after injection, the EEG had returned to baseline levels in the last two groups. About 6 days after SE, EEG was still of less amplitude than before injection of pilocarpine and in most groups peaks could still be recorded, occasionally in pilo-DZP, -TC10 and -TC30 mice. In the 78 pilo-TC60 rats, the frequency of high amplitude peaks was higher than in all other groups.

Following injection of test compound or vehicle, the EEG recording was not affected by injection into the pilo-DZP and pilo-TClO groups. In pilo-TC30 rats, the injection induced the occurrence of slow waves in hippocampal and cortex EEG and a lower peak frequency in pilo-TC60 rats.

All rats exposed to DZP, TC10 and TC30 and studied to the chronic phase developed SRS with a similar latency period. The latency period was 18.2 ± 6.9 days (n = 9) in pilo-DZP rats, 15.4 ± 5.1 days (n = 7) in pilo-TClO rats, 18.9 ± 9, 0 days (n = 10) in pilo-TC30 rats. In the group of rats submitted to TC60, one subgroup of rats became epileptic with a latency period similar to that of the other groups, i.e. and. 17.6 ± 8.7 days (n = 7) while another group of rats became epileptic with a much longer delay ranging from 109 to 191 post-SE days (149.8 ± 36.0 days, n = = 4) and a mouse did not become epileptic within 9 months after SE. The difference in latency period for SRS between pilo-DZP, pilo-TClO, pilo-TC30 and the first subgroup of pilo-TPM60 rats was not statistically significant. None of the saline-physiological serum (n = 5) mice developed SRS.

To calculate the frequency of SRS in rats exposed to pilocarpine, the severity of the seizure was measured and the phase III seizures (clonic convulsions of the facial and anterior limbs) and phase IV-V (steepness and fall) were distinguished. The frequency of phase III SRS per week in the pilo-DZP and pilo-compost assay rats was variable between the groups. It was low, constant in the pilo-DZP and pilo-TC60 groups (with early onset of SRS) during the first 3 weeks and disappeared during the 4th week in the pilo-DZP group. The frequency of phase III SRS was greater in the pilo-TClO group where it was significantly increased compared to the pilo-DZP values during weeks 3 and 4. The frequency of the most severe phase IV-V SRS was highest during the weeks in most groups except for the pilo-TC30 and TC60 with late seizure appearance where the SRS frequency was constant over the 4 weeks in the TC30 group and over the first two weeks in the pilo-TC60 group with late onset of SRS without IV-V phase seizures where no seizures were recorded after the second week. The frequency of phase IV-V SRS was significantly reduced in groups TC10, TC30 and TC60 (with early onset of SRS) (2.3-6.1 SRS per week) relative to the pilo-DZP group (11.3 SRS per week) during the first few weeks. During weeks 2-4v, the frequency of phase IV-V SRS was reduced in all groups over the first week, reaching 2-6 seizures per week, except in the pilo-TC60 group with early onset of SRS, where the frequency of seizures was significantly reduced to 0.6-0.9 seizures per week compared to the pilo-DZP group in which the SRS frequency ranged from 3.3 to 5.8.

Cellular densities in the hippocampus, thalamus and cortex

In pilo-DZP rats compared to saline-physiological serum rats, cell numbers were massively decreased in the hippocampal CAI region (70% cell loss in the pyramidal cell layer) while the CAS region was less extensively impaired (54% cell loss in CA3a and 31% in CA3b). In the dentate gyrus, pilo-DZP mice experienced extensive cell loss in the hilus (73%) while the granule cell layer showed no visible deterioration. Similar decay was observed in the ventral hippocampus but no cell counts were performed in this region. Extensive deterioration in the lateral thalamic nucleus (91% cell loss) was also recorded while the mediodorsal thalamic nucleus was more moderately damaged (56%). In the piriform cortex, cell loss was total in layers III-IV which was no longer visible and reached 53% in layer II in pilo-DZP rats. In the dorsal entorhinal cortex, layers II and III-IV suffered a slight deterioration (9 and 15%, respectively). The II layer of the ventral entorhinal cortex was fully preserved while the III-IV layers suffered a 44% cell loss.

In the hippocampus of pilo-test animals, the cell loss was reduced compared to pilo-DZP mice in the CAI pyramidal layer in which the cell loss reached 75% in pilo-DZP animals and 35 and 16% in pilo-TC30 animals or pilo-TC60, respectively. This difference was statistically significant at both doses of test compound. In the pyramidal layer CAS, the test compound did not provide any protection in the CA3a area while the 60 mg / kg dose of the test compound was significantly neuroprotective in CA3b. In the dentate gyrus, cell loss in hilus was similar in the pilo-test animals (69-72%) and pilo-DZP (73%). In both thalamic nuclei, the dose of 60 mg / kg was also protective by reducing the neuronal deterioration in 65 and 42% in the lateral and mediodorsal nuclei, respectively. In the cerebral cortex, treatment with the test compound provided neuronal protection relative to DZP alone at the higher dose, 60 mg / kg. At the two lowest doses, 10 and 30 mg / kg, the total cell loss and tissue disorganization observed in layers III-IV of the piriform cortex were identical in the pilo-DZP and pilo-test compound rats and allowed no counting in any of the groups. In layers II and III-IV of the piriform cortex, treatment with TC60 reduced the neuronal deterioration recorded in pilo-DZP rats by 41 and 44%, respectively. In the ventral entorhinal cortex, neuroprotection was induced by the administration of TC60 in layers III-IV and reached 31% by comparison with the pilo-DZP mice. In the entorhinal cortex, there was a slight worsening of cell loss in pilo-TClO rats relative to pilo-DZP rats in the III-IV layers of the dorsal entorhinal cortex (28% parents of deterioration) and in the III-IV layers of the ventral entorhinal cortex (35 % country of deterioration). At the other doses of the test compound, the cellular loss in the entorhinal cortex was similar to that recorded in the pilo-DZP mice.

Development of mossy fiber in the hippocampus

All rats presenting SRS in the pilo-DZP and pilo-TPM groups showed similar intensity of Timm development in the internal molecular layer of the dentate gyrus (ranks 2-4). Timm's revelation was present in the upper and lower blades of the dentate gyrus. The mean value of Timm index in the upper lamina reached 2.8 ± 0.8 in pilo-DZP rats (n = 9), 1.5 ± 0.6 in pilo-TClO rats (n = 7), 2.6 ± 1.0 in pilo-TC30 (n = 10) and 1.5 + 0.7 in the whole group of pilo-TC60 (n = 11) mice. When the pilo-compound test group at 60 mg / kg was subdivided according to the latency period up to SRS, the subgroup with early SRS had an 82

Timm index of 1.8 ± 0.6 (n = 6) and the subgroup of late-onset or non-SRS mice had a Timm index of 1.2 ± 0.6 (n = 5). The values recorded in the pilo-DZP rats were significantly different in statistical terms from the pilo-TClO values (p = 0.032) and the pilo-TC60 subgroup with late or absent seizures (p = 0.016).

Discussion and conclusions

The results of this study show that a 7-day treatment with the test compound starting 1 h after the onset of SE, is able to protect some areas of the brain from neuronal decay, e.g. in the pyramidal cell layer of the CAI and CA3b areas, the mediodorsal thalamus, the II and II MV layers of the pyriform cortex and the III-IV layers of the ventral entorhinal cortex, but only at the highest dose of the test compound, i. 60 mg / kg. The last dose of the test compound is also capable of delaying the occurrence of SRS in at least one subgroup of animals that became epileptic with an average delay which was about 9 times longer than in the other groups of animals and one non-susceptible animal. became epileptic within 9 months after SE.

These results show that a compound having anti-ictal properties, which are the classical properties of most of the antiepileptic drugs marketed, is also capable of retarding epileptogenesis, and. be antiepileptogenic. Data from this study also show that treatment with test compound, regardless of the dose used, decreases the severity of epilepsy as it decreases the number of IV-V phase seizures, especially during the first 83 weeks of the entire observation period of 4 weeks with treatment with the test compound at 60mg / kg. In addition, in the TC10 group, there is a shift towards an increase in the occurrence of less severe phase III seizures which are more numerous than in the pilo-DZP group. EXAMPLE 2 The aim of this extended part of the study was to continue the study described in Example 1 above relative to the potential neuroprotective and antiepileptogenic properties of the same Test Compound (TC) in the lithium-pilocarpine (Li-Pyl) model of wolf epilepsy temporal. In the first study it was shown that CT was able to protect the CAI and CA3 areas of the hippocampus, piriform cortex and ventral entorhinal cortex of neuronal deterioration induced by the Li-Pilo epileptic status. Most of these neuroprotective properties occurred at the highest dose studied, 60 mg / kg and the treatment was able to delay the occurrence of spontaneous seizures in 36% (4 of 11) of the rats. In this example the consequences of treatment with higher doses of CT in neuronal deterioration and epileptogenesis are studied. The lithium-pilocarpine model of temporal lobe epilepsy The rat-induced epilepsy model by lithium-associated pilocarpine (Li-Pyl) reproduces the majority of the clinical and neurophysiological specificities of human temporal lobe epilepsy (See, Turski L, Ionomonomé C, Turski WA, Bortolotto ZA, Cavalheiro EA (1989) Review: Cholinergic 84 mechanisms and epileptogenesis. intractable epilepsy, Synapse 3: 154-171, Cavalier EA (1995) The pilocarpine model of epilepsy, Ital J Neurol Sci., 16: 33-37).

In adult rats, systemic administration of pilocarpine leads to SE which may last up to 24 h. The lethality rate reaches 30-50% during the first few days. In surviving animals, neuronal deterioration predominates in hippocampal formation, in the piriform and entorhinal cortices, thalamus, amygdaloid complex, neocortex and substantia nigra. This period of acute convulsion is followed by a " silent " which persists for an average duration of 14-25 days after which all animals exhibit spontaneous convulsive seizures recurring at the usual frequency of 2 to 5 per week (See, Turski L, Ionomonomi C, Turski WA, Bortolotto ZA, Cavalheiro (1995) The pilocarpine model of epilepsy, Ital J Neurol Sci., 16: 33-33. [J Neurol Sci. 37, Dubé C, Boyet S, Marescaux C, Nehlig A (2001) Relationship between neuronal loss and interictal glucose metabolism during the chronic phase of the lithium-pilocarpine model of epilepsy in the immature and adult rat Exp Neurol 167: 227-241 )).

Current antiepileptic drugs (AEDs) do not prevent epileptogenesis and are only transiently effective in recurrent seizures.

In the applicant's previous study, we studied the potential neuroprotective and antiepileptogenic effects of increasing doses of Test Compound (TC) given as monotherapy and compared with our standard treatment of diazepam (DZP) administered primarily to prevent elevated mortality. These data show that a 7-day treatment with 10, 30 or 60 mg / kg TC starting 1 h after the onset of SE is able to protect some areas of the brain from neuronal deterioration. This effect is statistically significant in the pyramidal cell layer of the CAI and CA3b areas, in the mediodorsal thalamus, layers II and III-IV of the pyriform cortex and layers III-IV of the ventral entorhinal cortex, but only at the higher dose of CT, 60 mg / kg. In addition, it appears that the last dose of CT is also the only one capable of delaying the occurrence of SRS, at least in a subgroup of animals that became epileptic with an average delay that was about 9 times longer than in the other groups of animals and one animal did not become epileptic within 9 months after SE.

In this study, the effects of different doses of Test Compound (TC), i. 30, 60, 90 and 120 mg / kg (TC30, TC60, TC90 and TC120) using the same design as in the previous study. Treatment was started one hour after the onset of SE and the animals were treated with a second injection of the same dose of drug. This initial treatment of SE was followed by a 6-day CT treatment. This report refers to the effects of four different doses of CT on neuronal decay evaluated in the hippocampus, para-hippocampal cortices, thalamus and amygdala at 14 days after SE and in the latency period and frequency of spontaneous epileptic seizures. 86 Methods

Animals

Adult male Sprague-Dawley rats provided by Janvier Breeding Center (Le Genest-St-Isle, France) were housed under controlled current conditions with low animal density at 20-22 ° C (light / dark cycle, 7.00 am-7, 00 pm lights on), with food and water available ad libitum. All animal experiments were carried out in accordance with the rules of the Directive of the Council of the European Communities of 24 November 1986 (86/609 / EEC) and the French Department of Agriculture (License No. 67-97).

Induction of epileptic status, treatment with Test Compound (TC) and occurrence of SRS

All rats were given lithium chloride (3 meg / kg, ip, Sigma, St Louis, Mo, USA) and about 20 h later, all animals received also methylscopolamine bromide (1 mg / kg, sc, Sigma) was administered to restrict the peripheral effects of the seizure. SE was induced by injecting pilocarpine hydrochloride (25 mg / kg, s.c., Sigma) 30 min after methylscopolamine. The effects of increasing doses of CT were studied in 5 groups of rats. Animals received 2.5 mg / kg DZP, i.e., or 30, 60, 90 or 120 mg / kg TC (TC30, TC60, TC90, TC120), i.p., 1 h after the onset of SE. The control group received vehicle instead of pilocarpine and CT. Rats surviving SE were then injected, about 10 h after the first CT injection, with a second i.p. injection. of 1.25 mg / kg of DZP for the DZP group or with the same dose of TC 87

which in the morning and were maintained under treatment (s.c.) with TD twice daily for another 6 days, while DZP mice received a vehicle injection.

The effects of DZP and the 4 doses of TC on epileptogenesis were investigated by daily recording of the animals on video for 10 h per day. Video recording was performed for 4 weeks during which the first seizure was noted, as well as the total number of seizures throughout the period. The video recording system was then removed from the animals and these were kept for another 4 weeks in our animal facilities before being slaughtered after a total period of 8 weeks of epilepsy. Mice that did not exhibit seizures were sacrificed after 5 months of video recording.

Quantification of cell densities Quantification of cell densities was performed two times after SE: a first group was studied 14 days after SE and was composed of 7 DZP mice, 8 TC30 rats, 11 TC60 rats, 10 TC90 rats, 8 TC120 rats and 8 control mice not submitted to SE. A second group used to study the latency period up to SRS was sacrificed 8 weeks after the first SRS or at 5 months when SRS was no longer observable at that time and was composed of 14 DZP rats, 8 TC30 rats, 10 TC60 rats, 11 rats TC90 rats, 9 TC120 rats. At the moment neuronal counting is still taking place in the second group of animals studied for epileptogenesis and long-term counts and data for this part of the study will not be included in this report.

For neuronal count, the animals were deeply anesthetized with 1.8 g / kg pentobarbital (Dolethal®, Vétoquinol, Lure, France). The brains were removed and frozen. Successive slices of 20 μm were cut in a cryostat, air dried for several days prior to thionine development. Quantification of cell densities was performed with a microscopic grid of 1 cm2 with 10 x 10 boxes in coronal sections according to the stereotactic coordinates of the mouse brain atlas (Paxinos G, Watson C (1986) The Rat Brain in Stereotaxic Coordinates, 2nd ed Academic Press, San Diego). The counting grid was placed in a well-defined area of the brain structure of interest and counting was performed with a 200 or 400-fold microscope magnification defined for each single brain structure. Cell counts were performed twice on each side of three adjacent sections for each region by a single observer unaware of the treatment of the animal. The mean number of cells obtained in the 12 count fields in each brain structure was determined. This process was used to minimize potential errors that could result from double counts that would lead to an excess estimate of the number of cells. The neurons that touched the lower and right ends of the grid were not counted. The counts involved only neurons with cell bodies greater than 10 Âμm. Cells with smaller cell bodies were considered as glial cells and were not counted.

Data analysis

For neuronal deterioration and epileptogenesis, the statistical analysis between groups was performed using a unifactorial analysis of the variance, followed by a Dunnett or Fisher post-hoc test using Statistica software.

Results

Characteristics of the epileptic status of lithium-pilocarpine

A total number of 143 Sprague-Dawley rats weighing 250-330 g were subjected to lithium-pilocarpine-induced (Li-pyl) SE. Of this number, 10 did not develop SE while 133 mice developed a complete FE Li-pilo. The behavioral characteristics of SE were identical in the li-pilo-DZP and li-pilo-TC groups. Within less than 5 min after injection of pilocarpine, the rats developed diarrhea, piloerection and other signs of cholinergic stimulation. During the next 15-20 min the rats showed oscillating, shaving, chewing and exploratory behavior. Recurrent seizures began about 15-20 min after administration of pilocarpine. These seizures, which associated episodes of myoclonus of the head and bilateral of the hind limbs with steepness and fall, progressed to SE about 35-40 min after pilocarpine, as previously described (Turski L, Ionomonomouw C, Turski WA, Bortolotto ZA, Cavalheiro The relationship between neuronal loss and synaptic neuropathy in the treatment of epilepsy, as well as in the treatment of epilepsy, and interictal glucose metabolism during the chronic phase of the lithium-pilocarpine model of epilepsy in the immature and adult rat Exp Neurol 167: 227-241, André V, Rigoulot MA, Koning E, 90

Ferrandon A, Nehlig A (2003) Long-term pregabalin treatment protects basal cortices and delays the occurrence of spontaneous seizures in the lithium-pilocarpine model in the rat. Epilepsy 44: 893-903). The control group not submitted to SE and receiving lithium and saline was composed of 20 rats.

In the group of 57 cells counted at 14 days after SE, a total number of 13 mice died during the first 48 h after SE. Mortality varied with treatment: 36% (4/11) of DZP mice, 33% (4/12) of TC30 mice, 8% (1/12) of TC60 mice, 0% (0/10) of TC90 and 33% (4/12) TC120 mice. In the DZP group, the 4 rats died in the first 24 h after SE. In the TC30 rat group, one mouse died on SE day, one mouse was dead about 24 h after SE and two mice about 48 h. In the TC60 rat group, a mouse died at 48 h after SE. In the TC120 rat group, two mice were dead about 24 h and two about 48 h after SE.

In the group of 55 animals destined to study the latency period up to SRS and late cell count, the mortality rate during the first 48 h after SE was as follows: 7% (1/14) of DZP mice died, 27% ( 3/11) of the TC30, 0% (0/10) TC60 mice, 0% (0/11) TC90 mice and 0% (0/9) TC120 mice. In the DZP rat group, a mouse died during the first 24 h after SE. In the TC30 group, two mice were dead about 24 h and one about 48 h after SE. 91

Cellular densities in the hippocampus and cortex in the early phase (14 days after SE)

In DZP rats relative to control mice, the number of neurons was massively decreased in the hippocampal CAI region (85% decrease in the pyramidal cell layer) while the CA3 region was less extensively impaired (40% loss) (Table 1 and Figure 1) . In the dentate gyrus, DZP mice experienced extensive neuronal loss in the hilus (65%) while the granule cell layer showed no noticeable deterioration. The same distribution of decay in the ventral hippocampus was observed but cell counts were not performed in this region.

In the thalamus, neuronal loss was moderate in the central and lateral mediodorsal, in the dorsal medial dorsolateral and in the central medial nuclei (18, 24, 40 and 34% decrease, respectively), more pronounced in the mediodorsal nucleus (49%) and very important in the lateral ventral division of the dorsolateral nucleus (90%) (Table 1 and Figure 2). In the amygdala, the neuronal loss was moderate in the ventral medial posterior nucleus (38%) and more pronounced in the anterior basolateral and medial dorsal nuclei (73% and 53% decrease, respectively). There was no neuronal deterioration in the central nucleus (Table 1 and Figure 3).

In the piriform cortex, neuronal loss was almost complete in layer III (94%) which was no longer visible and reached 66 and 89% in the dorsal and ventral layer II, respectively, in the DZP rats relative to the saline-treated control rats. In the dorsal entorhinal cortex, layers II and III-IV suffered a slight deterioration (18 and 24%, respectively) and 92 in the ventral layers II and III / IV, the deterioration reached 22 and 74%, respectively (Table 1 below and Figure 4).

Table 1: Effects of increasing doses of Test Compound (TC) on the number of neuronal cell bodies in the hippocampus, thalamus, amygdala and cerebral cortex of rats submitted to IL-1β.

(N = 10) pilo-TC120 (n = 8) Hippocampus Area CAI 74 (n = 10) pilo-TC30 (n = 8) pilo-TC30 , 8 ± 1.5 10.9 ± 1.9 ** 39.3 ± 4 4 ** 00 31.9 ± 4 4 * * oo 47.7 ± 6.6 * 65.5 ± 2.09 ° Area CA3 52.1 ± 2.7 31.3 ± 2.9 9 ** 35.7 ± 1.8 ** 31.6 ± 1.4 ** 35.1 ± 2.9 ** 39.8 ± 1 , 5 ** Hilus 96.4 ± 3.5 33.5 ± 3.0 ** 33.0 ± 3.2 ** 32.8 ± 3.3 ** 37.5 ± 3.1 ** 44, 8 ± 2, 9 ** Medial Mediodorsal Thamo 31.9 ± 0.9 16.4 ± 1.9 ** 11.5 ± 2.5 ** 19.1 ± 2.6 ** 23.1 ± 2, 8 °° 28.6 ± 0.8 ° Central mediodorsal 31.9 ± 1.2 26.3 ± 1.8 ** 26.9 ± 0.6 * 24.1 ± 1 ** 27.4 ± 1 , 5 29.9 ± 1, 7th lateral mediodorsal 25.9 ± 0.6 19.6 ± 0.8 ** 20.5 ± 0.7 ** 18.9 ± 0.6 ** 22 ± 1.2 * 24.4 ± 1.1 ° Dorsolateral, medial, dorsal 102.2 ± 2.5 61 ± 6.3 ** 64, 2 ± 9.3 * * ° 77.5 ± 3 ^ 9 ** 00 79, 4 ± 3, 1 ** 89.8 ± 3.7 * Dorsolateral, lateral ventral 97.8 ± 1.7 9.7 ± 2: 5 ** 8.8 ± 2.8 ** 56.7 ± 8.7 ** 71.8 ± 5.3 ° * 79.0 ± 4, 7 ° C Median central 113.1 ± 5.9 74.2 ± 7.4 75.6 ± 7.7 83.7 ± 9.6 88.2 ± 8.5 108.2 ± 6 , 6 ° Basolateral Amygdala 46.7 ± 1.2 12.8 ± 5.3 ± 27.3 ± 4.9 ± 27.8 ± 4.3 ± 10 ± 40.7 ± 1.6 ° 42.7 ± 1.300 Medial, anterior dorsal 84.3 ± 3.8 40.0 ± 2.5 ** 46.8 ± 5.00 ** 58.4 ± 2.8 ** 72.2 ± 5, 7 ° ° 80.2 ± 2.6 °° Medial, ventral posterior 35.1 ± 1.7 21.8 ± 2.4 ** 22.3 ± 1.8 ** 26.2 ± 2, 9 ** 30, 7 ± 3, 7 °° 34.7 ± 1.7 ° Piriformis cerebral cortex, layer II, dorsal 36.6 ± 0.8 12.6 ± 4.2 ** 15.7 ± 2 , 9 ** 27.5 ± 2.8 ** ± 32.4 ± 1.1 ° ± 35.2 ± 1.1 ° Piriformis, layer II, ventral 33.0 ± 0.8 3.6 ± 0.7 ** 7.2 ± 3.8 ** 13.7 ± 4.2 ** 18.4 ± 4.0 0 ° 30.5 ± 1.3 °° 93 (continued)

(N = 10) pilo-TC90 (n = 10) pilo-TC30 (n = 8) pilo-TC30 (n = 8) pilo-TC60 19.2 ± 0.7 1.2 ± 1.2 ** 1.8 ± 1.8 ** 6.4 ± 2.3 ** 9 ± 3.0 °° 15 ± 2.2 °° Entorrinal, layer II, dorsal 29 ± 0.6 23.5 ± 0.7 ** 23.4 ± 0.6 ** 23.9 ± 0.5 ** 26.3 ± 0.9 ** 27.3 ± 0 , 5th ° Entorrinal, layer II, ventral 26.8 ± 0.7 21.7 ± 1.3 ** 22.7 ± 0.9 23.3 ± 0.8 ** 25.4 ± 1.1 ° 25.1 ± 0.6 Entorrinal, layer III / IV, dorsal 29.2 ± 0.9 22.3 ± 0.5 ** 22.3 ± 0.5 ** 23.2 ± 0.8 ** 26 , 7 ± 0.8 * 26.4 ± 0.70 ° C. Entorrinal, layer III / IV, ventral 28.7 ± 1.7 7.7 ± 2.3 ** 13.2 ± 1.9 ** 16 , 5 ± 2.2 ** 23.7 ± 1.5 °° 24.5 ± 1.4 °° * p < 0.05, ** p &lt; 0.01, statistically significant difference between the pilo-S TC and physiological serum-control rats p < 0.05, °° p &lt; 0.01, statistically significant difference between the pilo-S TC and pilo-DZP \

In the hippocampus of CT treated animals, the cell loss was significantly reduced relative to DZP rats in the CAI pyramidal cell layer. This reduction was accentuated in the TC30, 60 or 90 (36-47% cell loss) and prominent TC120 (12% cell loss) rats. The differences were statistically significant at all TC doses (Table 1 and Figure 1). In the pyramidal layer CA3, there was a tendency for slight neuroprotection induced by the Test Compound, only at the dose of 120 mg / kg but the difference with the DZP group was not significant. In the dentate gyrus, cell loss in hilus was similar in the DZP and TC30, 60 and 90 (61-66% decrease) groups and there was a slight tendency for a lower deterioration in the TC120 group (53% of neuronal loss) compared to the animals DZP (66% decrease). None of these differences was statistically significant.

In the thalamus, neuronal loss was similar in DZP and TC30 and TC60 rats. CT was significantly protective at the dose of 94 60 mg / kg in the dorsal medial dorsolateral nucleus and at two higher doses, 90 and 120 mg / kg in all thalamic nuclei, although the difference was not significant in the central mediodorsal and medial central nuclei in TC90 rats. In TC120 mice, neuronal decrease was considerably reduced relative to DZP mice. It ranged from 4-19% and the number of neurons was no longer significantly different from control animals, except in the dorsal medial dorsolateral nucleus (Table 1 and Figure 2). In the amygdala, CT was significantly protective at the 30 mg / kg dose in the basolateral nucleus and at the 60 mg dose, also in the anterior medial dorsal nucleus. At the highest dose, CT was largely neuroprotective; the number of neurons ceased to be significantly different from the control level and reached 86-99% of the control level in all amygdala nuclei (Table 1 and Figure 3).

In the cerebral cortex, CT treatment did not significantly protect any cortical area relative to DZP treatment at a dose of 30 mg / kg. At 60 mg / kg, CT significantly reduced neuronal loss only in layer II of the dorsal piriform cortex (25% decrease compared to 66% in the DZP group) At 90 and 120 mg / kg, TC significantly protected all three areas of the piriform cortex compared to treatment with DZP and at the highest dose of TC, 120 mg / kg, neuronal density reached 78-96% of control levels, even in layer II and layer III of the dorsal piriform cortex where the neuronal population was almost completely eliminated in the DZP group. In all layers of the dorsal and ventral entorhinal cortex, the two lower doses of CT, 30 and 60 mg / kg provided no neuroprotection. The 90-mg / kg dose of TC significantly protected layers II and III / IV of the ventral entorhinal cortex (4 and 17% deterioration remaining in the 95 layers II and II / IV of the dorsal part and layer II of the ventral part in compared with 19 and 73% in the DZP group). At the highest CT dose, 120 mg / kg, all parts of the entorhinal cortex, both dorsal and ventral cortex were protected and the number of neurons in these areas ceased to be significantly different from the level in the controls (85-94% of neurons survivors compared with 27-81% in the DZP group). The latency period and the frequency of recurrent seizures

The latency period for spontaneous seizures reached a mean value of 15.5 ± 2.3 days in the DZP group (14 rats) and was similar (11.6 ± 2.5 days) in the TC30 group (8 rats). At higher CT concentrations, the animals could be subdivided into subgroups with short and long latency periods. It was considered a short latency period as any duration shorter than 40 days after SE. Some mice exhibited a latency period for the first spontaneous seizure which was similar to that recorded in the DZP and TC groups but the number of mice exhibiting these short latency values decreased progressively with increasing TC concentration. Thus, at 30 mg / kg, 70% of the rats (7/10) had short latency times for seizures while at 90 and 120 mg / kg, this percentage reached 36% (4/11) and 11% (1/9 ), respectively (Table 2 below and Figure 5). 96

Table 2: Effect of increasing doses of CT in the latency period for spontaneous seizures.

(Number of days) DZP 14 15.5 ± 2.34 pilo-TC30 8 11.6 ± 2.5 pilo-TC60 10 2 groups Short latency period (n = 7) Long latency period (n = 3) 17.4 ± 5.4 76.7 ± 15.6 ** ° pilo-TC90 11 3 groups Short latency period (n = 4) Long latency period (n = 2) ) Non-epileptic (n = 5) 14.8 ± 5.7 52.0 ± 1.0 * 150 ** 00 3 groups pilo-TC12 0 9 Short latency period (n = 1) Long latency period = 4) Non epileptic (n = 4) 13.0 84.5 ± 16.7 ** 0 0 150 ** 00

** p &lt; 0.01, * p &lt; 0.05, statistically significant differences compared to the pito-DZP 00 p &lt; 0.01, ° p &lt; 0.05, statistically significant differences compared to the short latency period group

In groups TC60, 90 and 120, the mean value of the rats with long latency periods was similar and ranged from 52 to 85 days. Finally, at two higher doses of CT, we were able to identify a percentage of mice that did not develop any seizures over a period of 150 days post SE. The percentage of non-epileptic rats reached 45% at both doses of TC. The frequency of spontaneous seizures was similar over the four weeks of enrollment. It showed a tendency to be higher in the DZP and TC30 groups, while it was lower in the TC60, 90 and 120 groups (Figure 6). These differences did not reach statistical significance at the level of each individual weekly frequency but reached significance for the total or mean number of seizures over four weeks. The number of seizures was also plotted according to the duration of the latency period for the first spontaneous seizure. Animals with a short latency period showed a tendency to present 2-3 times more seizures over the four weeks of recording than mice with a long latency period. No statistical analysis could be performed since ANOVA showed no significance, most likely because there was only one animal in the short-latency subgroup of TC120 animals (Figure 7). However, when all latency periods were plotted against the number of seizures, there was a significant inverse correlation that gave rise to a straight line with a correlation coefficient of -0.4 (Figure 8).

To complete this analysis two more measurements will be taken. The first is the cell count in animals that were recorded on video and followed for 2 months after the first spontaneous seizure or sacrificed at 5 months to study the potential correlation between the extent and location of brain deterioration and the occurrence and / or period of latency for spontaneous seizures. The second will be to perform a one-year follow-up on the occurrence of seizures in a group of rats to study whether the animals the applicant considered "non-epileptic &quot; at 5 months remain free of seizures or not.

The results of the present study show that a CT treatment started 1 h after the onset of SE induced by 98

Li-pilo has neuroprotective properties in the CAI pyramidal cell layer of the hippocampus and in all layers of the ventral and dorsal and entorhinal pyriform cortex. CT also protects the nuclei of the thalamus and amygdala. However, CT is not protective at the dose of 30 mg / kg except for CAI, a thalamic nucleus and a tonsil nucleus. At the dose of 60 mg / kg, layer II of the dorsal piriform cortex and a second nucleus of the amygdala are also protected. At 90 and 120 mg / kg, the drug protects most of the brain regions studied except hippocampal CA3 and dentate gyrus hilus. The last two structures and the dorsal ventral dorsolateral thalamic nucleus are the only regions where the number of neurons remains significantly different from controls at the 120 mg / kg TC dose. From this data the extremely powerful neuroprotection properties of CT seem evident. The molecule appears to prevent neuronal death in most regions belonging to the Li-pilo-induced limbic epilepsy circuit, i. e., the hippocampus, thalamus, amygdala and para-hippocampal cortices. These are all regions in which we have detected MRI signals in the course of epileptogenesis in Li-pilo treated rats (Roch C, Leroy C, Nehlig A, Namer IJ (2002a) lithium-pilocarpine model of temporal lobe epilepsy in adult rats Epilepsy 43: 325-335). The only two regions that are not effectively protected by CT are the CA3 pyramidal cell layer and the hilus of the dentate gyrus. The latter region undergoes rapid and massive cell deterioration (André V, Marescaux C, Nehlig A, Fritschy JM (2001)). Hippocampus (GABAergic system contributes to the development of spontaneous recurrent seizures in the lithium-pilocarpine model of temporal lobe epilepsy. 11: 452-468. Roch C, Leroy C, Nehlig A, Namer IJ (2002a) Contribution of magnetic resonance imaging to the study of the lithium-pilocarpine model 99 of temporal lobe epilepsy in adult rats Epilepsy 43: 325-335 ) and none of the neuroprotection that the applicant has used in previous studies has been able to protect this structure. Based on previous studies, the applicant has also identified this structure as a key area in the initiation and maintenance of epileptic seizures in the Li-pilo model. Epilepsia 41 (Supl 6): S36-S43). The present study was carried out in the United States of America,

Obviously, current data demonstrate that epileptogenesis can be prevented even though deterioration remains very marked in this area. The long-term cell count in the group of animals that were recorded on video will be able to demonstrate whether the extent of deterioration in this region is critical for epileptogenesis in this model or not. The treatment did not affect the latency period for the first spontaneous seizure at the 30 mg / kg dose. At 3 higher doses, a percentage of animals developed epilepsy as rapidly as DZP or TC30 mice but the relative importance of this subgroup was inversely related to the dose of CT used. Another subgroup, constant in size (2-4 animals per group) developed epilepsy after a latency period 4-6 times longer, while at the two highest doses of the drug, 4-5 rats did not become epileptic after 5 months, i . e., about 10 times the duration of the short latency period and 2-3 times that of the long latency period. This delay in the occurrence of epilepsy may correlate with the number of neurons protected in the basal cortices in the animals. This assumption is based on the fact that the applicant has found some heterogeneity in the extent of neuroprotection in the basal cortices of animals subjected to short-term neuronal count at 14 days after SE. However, at the time, the remanent did not perform the neuronal count in the animals used for the study of epileptogenesis and therefore no conclusion can be drawn as to the potential relation between the number of surviving neurons in the basal cortices and the rate or even the occurrence of epileptogenesis.

The data obtained in the present study are in line with the earlier study in this group reporting that the 60 mg / kg dose of Compound Assay (CT) protected the hippocampus and basal cortices from neuronal deterioration and delayed the occurrence of recurrent seizures ( see Example 1). These confirm that protection of basal cortices could be a key factor in inducing a disease modifying effect on the lithium-pilocarpine model of epilepsy. The key role of the basal cortices as initiators of the epileptic process was previously demonstrated by the applicant's group in the lithium-pilocarpine model (André V, Rigoulot MA, Koning E, Ferrandon A, Nehlig A (2003) Long-term pregabalin treatment protects basal cortices and delays the occurrence of spontaneous seizures in the lithium-pilocarpine model in the rat Epilepsy 44: 893-903 Roch C, Leroy C, Nehlig A, Namer IJ (2002a) Contribution of magnetic resonance imaging to the study of the lithium- (pb-1), p-day-old rats (p21-day-old rats), and pituitary epilepsy (p21-day-old rats) : a MRI approach using the lithium-pilocarpine model Epilepsy 43: 1129-1136.

References to Example 2

André V, Marescaux C, Nehlig A, Fritschy JM (2001) Alterations of the hippocampal GABAergic System contribute to the development of spontaneous recurrent seizures in the lithium-pilocarpine model of temporal lobe epilepsy. Hippocampus 11: 452-468.

André V, Rigoulot MA, Koning E, Ferrandon A, Nehlig A (2003) Long-term pregabalin treatment protects basal cortices and delays the occurrence of spontaneous seizures in the lithium-pilocarpine model in the rat. Epilepsy 44: 893-903.

Cavalheiro EA (1995) The pilocarpine model of epilepsy. Ital J Neurol Sci., 16: 33-37.

Dubé C, Marescaux C, Nehlig A (2000) A metabolic and neuropathological approach to the understanding of plastic changes occurring in the immature and adult rat brain during lithium-pilocarpine induced epileptogenesis. Epilepsy 41 (Suppl 6): S36-S43.

Dubé C, Boyet S, Marescaux C, Nehlig A (2001) Relationship between neuronal loss and interictal glucose metabolism during the chronic phase of the lithium-pilocarpine model of epilepsy in the immature and adult rat. Exp Neurol 167: 227-241.

Paxinos G, Watson C (1986) The Rat Brain in Stereotaxic Coordinates, 2nd ed. Academic Press, San Diego.

Roch C, Leroy C, Nehlig A, Namer IJ (2002a) Contribution of magnetic resonance imaging to the study of the lithium-pilocarpine model of temporal lobe epilepsy in adult rats. Epilepsy 43: 325-335. 102

Roch C, Leroy C, Nehlig A, Namer IJ (2002b) Predictive value of cortical injury for the development of temporal lobe epilepsy in P21-day-old rats: a MRI approach using the lithium-pilocarpine model. Epilepsy 43: 1129-1136.

Turski L, Ikonomidou C, Turski WA, Bortolotto ZA,

Gentleman EA (1989) Review: Cholinergic mechanisms and epileptogenesis. The seizures induced by pilocarpine: a novel experimental model of intractable epilepsy. Synapse 3: 154-171. The Test Compound (TC) referred to in the example below is the compound of Formula 7 and the same compound as in the other examples 1 and 2 above. EXAMPLE 3 The aim of this study was to evaluate the pharmacokinetics (PK) of Test Compound (TC) following single oral administration and repeated in a healthy adult male at clinically relevant doses METHODS:

Two single-dose, double-blind, double-blind, placebo-controlled studies were conducted in healthy men aged> 18 and> 45 years. In study 1 (N = 70), subjects were randomly assigned to a single dose of Test Compound (TC) or placebo. Ascending doses such as 100, 250, 400, 750, 1000, 1250 and 1500 mg were received. PK parameters were estimated from plasma and urine samples collected up to 103 days after dose. Study 2 (N = 53) evaluated the PK of repeated doses of Test Compound (TC) in groups of 4 doses (100, 250, 500 or 750 mg). Within each group, 12 subjects were given q1h treatment with either drug or placebo for 1 week and were crossed after a 14 day elimination period. PK parameters were estimated from plasma and urine samples on days 1 and 7. RESULTS:

Single dose: The test compound (TC) was rapidly absorbed after oral administration. Cmax and AUC0 aumentar increased in proportion to the dose over the range of 100-1500 mg. The mean t max varied from 1.3-2.7 h. The mean values of ti / 2 (11.5-13.9 h), CUF (2.87-3.67 Uh) and Vd / F (52.1-66.3 Uh) were similar for all 7 groups of dose.

Repeated Doses: Plasma Test Compound (TC) concentrations reached steady state after 3-4 days as expected from their single dose half-life. The mean tmax occurred 1.3-1.8 h after administration. The mean values of 11/2 (11.9-12.8 h) and CUF (3.40-3.78 Uh) at steady state were comparable to PK parameters after single dose administration at day 1 and study 1 Cmax and AUC0-12 steady state increased in proportion to the dose.

As expected, there was a moderate degree of accumulation of Test Compound (T-C); Cmax and AUCo-12 were about twice as high on day 7 vs. day 1 (P &lt; 0.001). The mean CLr estimates for Test Compound (TC) were &lt; 5% of the mean oral clearance, suggesting non-renal clearance as the primary mechanism for the elimination of Test Compound (TC). CONCLUSIONS: Test Compound (TC) exhibited linear PK after single (100-1500 mg) and repeated doses (100-750 mg bid). It was rapidly absorbed and had a mean elimination half-life of 11.5-13.9 h after bid administration. Following gl 2 h administration, the Test Compound (CT) doubled up and was eliminated primarily by a non-renal pathway. EXAMPLE 4

Predictive example of a treatment regimen with the Test Compound (TC):

A 23-year-old male soldier is admitted to the hospital with a piercing wound on the head of a bomb fragment. The fragment entered the right frontal lobe of the brain and penetrated approximately one inch into the brain. The fragment is surgically removed and the patient recovers well with minimal neurological dysfunction. The patient has no personal or family history of a seizure disorder and shows no evidence of a seizure disorder after surgery and EEGs are negative in relation to seizure activity. The patient's physician immediately starts a treatment regimen with the Test Compound (TC) at a dose of 500 mg twice daily to prevent the development of a seizure disorder as a consequence of the injury. The prophylactic treatment is continued for one year and in the follow-up the patient shows no evidence of developing a seizure disorder. The patient is followed for another year and shows no evidence of developing a seizure disorder. EXAMPLE 5

Predictive example of a treatment regimen with the

Test Compound (TC):

A 3-year-old woman is admitted to the hospital after suffering from blunt head injury in a car accident. The patient has no past of any type of convulsive disorder and the personal and family clinical past are essentially negative. The patient's head hit the dashboard of the car and it lost consciousness for 30 minutes after the accident. CT scanning shows a small contusion in the left frontal region of the brain and the patient is diagnosed as having blunt head trauma with a concussion. The patient's physician is concerned about the possible future development of a seizure disorder as a result of the injury to the patient's frontal lobe. The patient's physician immediately initiates a treatment regimen with Test Compound (TC) at a dose of 300 mg twice daily to prevent the development of a seizure disorder. The patient is followed for a year and shows no evidence of seizure development. The dose of Test Compound is gradually reduced and terminated and the patient remains seizure free at follow-up two years later. EXAMPLE 6

Predictive example of a treatment regimen with the Test Compound (TC):

A 74-year-old woman is admitted to the hospital after an episode at home in which she developed weakness on the right side and inability to speak. An MRI showed a heart attack of the left middle cerebral artery. The patient had no personal or family history of seizures or other neurological problems. In addition to routine supportive care, the patient's physician initiated a Test Compound (CT) regimen at a dose of 250 mg twice daily to prevent the development of a seizure disorder in the post-heart attack recovery period. The patient is seen at a follow-up visit a year later and has no evidence of developing a seizure disorder and the medication is gradually reduced. The patient continues to be free from convulsions following the end of two years. EXAMPLE 7

Predictive example of a treatment regimen with the Test Compound (TC):

A previously 7-year-old boy is examined in his doctor's office after having repeated seizures at home in the context of a 105-degree fever secondary to a viral infection of the upper respiratory tract. The patient has recovered from viral infection and shows no signs of having a seizure disorder. The physician 107 diagnoses febrile seizures. The boy weighs 27 kilograms and the doctor decides to initiate a Test Compound (CT) regimen at a dose of 7.1 mg / kg / day and, therefore, the boy is given 100 mg twice a day to prevent development of a seizure disorder. The patient is seen at the follow-up appointment after one year and is free of seizures. The medication is continued for two years and finished. The patient does not show any evidence of a seizure disorder, followed up for three years. EXAMPLE 8

Predictive example of a treatment regimen with the Test Compound (TC):

A 47-year-old man is admitted to the hospital for excision of a right pre-frontal AVM. The patient does not have any personal or family history of seizure disorder. Before the neurosurgical procedure, the patient's physician initiates a Test Compound regimen at a dose of 200 mg twice daily to minimize the risk of developing a seizure disorder after surgery. The patient is seen at the follow-up consultation one year after the procedure and the medication is reduced and terminated. The patient remains well and shows no evidence of developing a seizure disorder in the follow-up three years later. The present invention is not limited by the particular embodiments or examples described in this application, which are mere illustrations of individual aspects of the invention. Many modifications and variations of this invention may be made without departing from its scope, as will be apparent to those skilled in the art. Functionally equivalent methods and combinations within the scope of the invention, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing description and examples and the accompanying drawings. Such modifications and variations fall within the scope of the appended claims. The present invention is only limited by the terms of the appended claims.

Lisbon, April 30, 2010 109

Claims (36)

A compound, or a pharmaceutically acceptable salt or ester thereof, selected from the group consisting of Formula (I) and Formula (II): wherein phenyl is X-substituted with one to five halogen atoms selected from the group consisting of fluoro, chloro, bromo and iodo; and, R1, R2, R3, R4, R5 and R6 are independently selected from the group consisting of hydrogen and C1 -C4 alkyl; wherein C 1 -C 4 alkyl is optionally substituted with phenyl (wherein phenyl is optionally substituted with substituents independently selected from the group consisting of halogen, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, amino, nitro and cyano) for use in the prevention, reversion, arrest or inhibition of epileptogenesis in a human subject who is at risk for developing epilepsy or a seizure-related disorder but has no epilepsy or clinical evidence of seizures. A compound as claimed in claim 1, wherein X is chloro. A compound as claimed in claim 1, wherein X is substituted at the ortho position of the phenyl ring. A compound as claimed in claim 1, wherein R1, R2, R3, R4, R5 and R8 are selected from hydrogen. An enantiomer or a pharmaceutically acceptable salt or ester thereof selected from the group consisting of Formula (I) and Formula (II) or enantiomeric mixture wherein an enantiomer selected from the group consisting of Formula (I) and Formula (II) predominates; Formula (II) wherein 2-phenyl is substituted on X with one to five halogen atoms selected from the group consisting of fluoro, chloro, bromo and iodo; and R 1, R 2, R 3, R 4, R 5 and R 6 are independently selected from the group consisting of hydrogen and C 1 -C 4 -alkyl; wherein C 1 -C 4 alkyl is optionally substituted with phenyl (wherein phenyl is optionally substituted with substituents independently selected from the group consisting of halogen, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, amino, nitro and cyano) for use in the prevention, reversion, arrest or inhibition of epileptogenesis in a human subject who is at risk of developing epilepsy or a seizure-related disorder but who does not have epilepsy or clinical evidence of seizures. An enantiomer as claimed in claim 5, wherein X is chloro. An enantiomer as claimed in claim 5, wherein X is substituted at the ortho position of the phenyl ring. An enantiomer as claimed in claim 5, wherein R1, R2, R3, R4, R5 and R6 are selected from hydrogen. An enantiomer as claimed in claim 5, wherein an enantiomer selected from the group consisting of Formula (I) and Formula (II) predominates in a content of from about 90% or more. 3 An enantiomer as claimed in claim 5, wherein an enantiomer selected from the group consisting of Formula (I) and Formula (II) predominates in a content of from about 98% or more. An enantiomer as claimed in claim 5, wherein the enantiomer selected from the group consisting of Formula (I) and Formula (II) is an enantiomer selected from the group consisting of Formula (Ia) and Formula (IIa): wherein phenyl is X-substituted with one to five halogen atoms selected from the group consisting of fluoro, chloro, bromo and iodo; and R 2, R 3 and R 4 are independently selected from the group consisting of hydrogen and C 1 -C 4 alkyl; wherein C1 -C4 alkyl is optionally substituted with phenyl (wherein phenyl is optionally substituted with substituents independently selected from the group consisting of halogen, C1 -C4 alkyl, C1 -C4 alkoxy, amino, nitro and cyano). An enantiomer as claimed in claim 11, wherein X is chloro. An enantiomer as claimed in claim 11, wherein X is substituted at the ortho position of the phenyl ring. An enantiomer as claimed in claim 11, wherein R1, R2, R3, R4, R5 and R8 are selected from hydrogen. An enantiomer as claimed in claim 11, wherein an enantiomer selected from the group consisting of Formula (Ia) and Formula (11a) predominates at a content of from about 90% or more. An enantiomer as claimed in claim 11, wherein an enantiomer selected from the group consisting of Formula (Ia) and Formula (IIa) predominates in a content of from about 98% or more. An enantiomer as claimed in claim 5, wherein the enantiomer selected from the group consisting of Formula (I) and Formula (II) is an enantiomer selected from the group consisting of Formula (Ib) and Formula (IIb) or a salt form thereof or a pharmaceutically acceptable ester: Formula (Ib) Formula (IIb) An enantiomer as claimed in claim 17, wherein an enantiomer selected from the group consisting of Formula (Ib) and Formula (llb) predominates at a content of from about 90% or more. An enantiomer as claimed in claim 17, wherein an enantiomer selected from the group consisting of Formula (Ib) and Formula (llb) predominates at a content of from about 98% or more. A compound or enantiomer as claimed in claims 1 or 5, wherein the predisposing factor (s) rendering the patient at risk for developing epilepsy or a seizure related disorder is selected from the group consisting of: injury or trauma of any type of CNS; neurosurgical procedures, activities at risk of CNS injury, e.g. combat activities, car or horse racing, and contact sports including boxing; spinal cord injury; infections of the CNS; anoxia; heart attack (CVA); past Transient Ischemic Attacks (TIA); carotid stenosis; past disease of atherosclerotic vessels; pulmonary embolism; peripheral vascular disease; Autoimmune diseases affecting the CNS, e.g. lupus; labor injuries, e.g. perinatal asphyxia; heart attack; therapeutic or diagnostic vascular surgical procedures, e.g. carotid endarterectomy or cerebral angiography; hypotension; CNS injury by embolisms, hyper- or hypoperfusion; hypoxia; known genetic predisposition for disorders known to respond to AEGD; CNS compression lesions; brain tumors, e.g. g. , glioblastomas; bleeding or bleeding in or surrounding the CNS, e.g. intracerebral bleeds or subdural hematomas; cerebral edema; febrile seizures; hyperthermia; exposure to toxic or poisonous agents; substance intoxication or withdrawal, e.g. g. cocaine, methamphetamine or alcohol; a family history of seizure disorders or a seizure-related neurological disorder related to epilepsy or seizure-related disorder, past seizure disorder; current treatment with medications that reduce the seizure threshold, e.g. g. , lithium carbonate, thorazine or clozapine; evidence from substitute markers or biomarkers that the patient is in need of treatment with an antiepileptogenic drug, e.g. g. MRI scan showing hippocampal sclerosis, elevated levels of serum neuronal degradation products, elevated levels of ciliary neurotrophic factor (CNTF), or an EEG suggestive of a seizure disorder or epilepsy-related seizure disorder. A compound or enantiomer as claimed in claim 20, wherein the predisposing factor (s) rendering the patient at risk of developing epilepsy or a seizure related disorder is selected from the group consisting of: closed head trauma or penetrating; neurosurgical procedures, carotid stenosis, heart attack or other stroke (CVA); epileptic status and CNS compression lesions. A compound or enantiomer as claimed in claim 21, wherein said predisposing factor (s) are closed head trauma or penetrating head trauma or a neurosurgical procedure. A compound or enantiomer as claimed in claim 21, wherein said predisposing factor (s) are heart attack, another cerebrovascular accident (CVA), presence of carotid stenosis or Transient Ischemic Attack. A compound or enantiomer as claimed in claim 23, wherein said predisposition factor is the epileptic state. 25. Compounds or enantiomers as claimed in claims 1 or 5, wherein said compound (or enantiomer) or a pharmaceutically acceptable salt or ester thereof is administered, in combination with one or more other compounds or therapeutic agents. A compound or enantiomer as claimed in claim 25, wherein said one or more other therapeutic compounds or agents are selected from the group consisting of compounds having one or more of the following: antioxidant activity; NMDA receptor antagonism; ability to increase inhibition of endogenous GABA; NO synthase inhibitory activity; iron fixing capacity, e.g. an iron chelating agent; calcium fixing capacity, e.g. a Ca (II) chelating agent; zinc fastening capacity, e.g. a Zn (II) chelating agent; the ability to block sodium or calcium ion channels; the ability to open the channels of potassium or chloride ions, therapeutic agents useful in the treatment of Substance Abuse. A compound or enantiomer as claimed in claim 25, wherein said one or more compounds are selected from the group consisting of anti-epileptic drugs (AED). A compound or enantiomer as claimed in claim 27, wherein said antiepileptic drug (AED) is selected from the group consisting of; carbamazepine, clobazam, clonazepam, etosuximide, felbamate, gabapentin, lamothiazine, levetiracetam, oxcarbazepine, phenobarbital, phenytoin, pregabalin, primidone, retigabine, talampanel, tiagabine, topiramate, valproate, vigabatrin, zonisamide, benzodiazepines, barbiturates or sedative hypnotics. A compound or enantiomer as claimed in claims 1 or 5, wherein the therapeutically effective amount is from about 5.7 mg / kg / day to about 42.9 mg / kg / day. A compound or enantiomer as claimed in claims 1 or 5, wherein said therapeutic amount is progressively decreased as the treatment of the epileptogenic process in said patient progresses. A compound or enantiomer as claimed in claims 25, 26, 27 or 28, wherein the amount of said one or more other compounds or therapeutic agents administered in association with said compound (or enantiomer) or a pharmaceutically acceptable salt or ester thereof is progressively diminished as the treatment of the epileptogenic process in said patient progresses. A compound or enantiomer as claimed in claims 1 or 5 wherein the administered dose is from about 6.4 mg / kg / day to about 35.7 mg / kg / day. A compound or enantiomer as claimed in claims 1 or 5 wherein the administered dose is from about 7.1 mg / kg / day to about 28.6 mg / kg / day. A compound or enantiomer as claimed in claims 1 or 5 wherein the dose administered is from about 7.9 mg / kg / day to about 21.4 mg / kg / day. A compound or enantiomer as claimed in claims 1 or 5 wherein the dose administered is from about 8.6 mg / kg / day to about 17.1 mg / kg / day. A compound or enantiomer as claimed in claims 1 or 5, wherein the dose administered is from about 400 mg / day to about 3000 mg / day. Enantiomer 38. 39. 40. Compound or claims 1 or 5, wherein about 450 mg / day to about Compound or enantiomer claims 1 or 5, wherein about 500 mg / day to about Compound or enantiomer claims 1 or 5, wherein about 550 mg / day to about Compound or enantiomer claims 1 or 5, wherein about 600 mg / day to about as claimed in the administered dose is from 2500 mg / day. as claimed in the administered dose is from 2000 mg / day. as claimed in the administered dose is from 1500 mg / day. as claimed in the administered dose is from 1200 mg / day. Lisbon, April 30, 2010 11